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Introduction

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Introduction

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1. INTRODUCTION

Every year millions of individuals diet to lose weight, but the long term prognosis

of such treatment is poor, since, to quote Albert Stunkard: Most obese persons will not stay

in treatment for obesity. Of those who stay in treatment, most will not lose weight and of

those who do lose weight, most will regain it (Stunkard, 1958). According to WHO (2010)

and Smith (2011), obesity is an excess of body fat frequently resulting in a significant

impairment of health and longevity. Obesity and overweight are caused by a chronic

imbalance between energy intake and energy expenditure. High intake of dietary fat, poor

exercise and sedentary lifestyle are the main causes for obesity. Obesity has several

adverse health effects such as hypertension, diabetes mellitus, hypercholesterolemia

and can even lead to cardiovascular disease (CVD) (Gensini et al., 1998; WHO, 2000;

Smith, 2011). The strong association between obesity and cancer has only recently come to

light (WHO, 2009).

The worldwide prevalence of obesity in adults is currently estimated to exceed

300 million (Cheetham et al., 2004). Although altering eating and activity behavior,

lifestyle is the cornerstone of anti-obesity, the pharmacotherapy for obesity is necessary.

However, most of the medications listed by United States food and drug administration

(FDA) were only approved for short-term use because patient’s complications to drug

therapy usually were exhibited (Karalik and Reilz, 1999; Cheetham et al., 2004). One of

the novel strategies for anti-obesity is to exploit the natural products from traditional

medicinal plants in the form of plant extracts or functional food. However, investigations

of pharmacological and action mechanisms of natural compounds and oriental remedies

were limited (Sachdewa and Khemani, 2003; Kim, et al., 2006; Lenon, 2012).

Obesity therapies include reducing nutrient absorption, applying anorectic and

thermogenic drugs that affect lipid mobilization and utilization. With the exception of

orlistat, a recently approved gastrointestinal lipase inhibitor, all drugs approved for the

treatment of obesity are either catecholaminergic or serotonergic CNS-active (activating

the sympathetic nervous system) anorectic agents (Van der Ploeg, 2000). Sibutramine is a

selective inhibitor of the reuptake of monoamines; primarily serotonin and noradrenaline

and to a lesser extent dopamine (Heal et al., 1998; Arterburn et al., 2004).

Upon termination of therapy with these drugs, weight is rapidly regained in many cases


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(Van der Ploeg, 2000). Because of the adverse effects associated with these anti-obesity

drugs, many trials have been recently conducted to find and develop new anti-obesity

drugs through herbal medicines that would minimize the side effects. Numerous animal

studies and clinical studies with various herbal medicines have been performed and some

studies reported significant improvements in controlling body weight without any

noticeable adverse effects (Yoshida et al., 1995; Heymsfield et al., 1998; Xie et al., 2002).

Some herbs and supplements such as 5-HTP, fiber and green tea may aid in weight

loss and help to alleviate the complications of obesity. Indian traditional medicinal plants

such as Commiphora weghtii (Guggulu), Allium cepa (Onion), Zingiber officinale

(Ginger), Trigonella foenum greecum (Fenugreek), Garcinia cambogia (Garcinia),

Gymnema sylvetra (Meshashringi), Terminalia chebula (Chebulic myrobalan, Haritaki),

Terminalia bellirica, Phyllanthus emblica, Plantego psyllium (Psyllium) and

Oenothera biennis (Primrose) have gained great reputation to reduce harmful cholesterol

and triglycerides in blood, prevent fat formation and reduce obesity (Sharma et al., 1990;

Srikumar et al., 2005). So, present study selected herbal drugs geriforte and haritaki for

evaluation of anti-obesity and hypolipidemic activity by using high fat diet induced obesity

in male albino rats.

1.1. LITERATURE PERTINENT TO OBESITY

Obesity has emerged as the most prevalent serious public health problem of our time

(Roberts and Mayer, 2000). It is a complex disorder, which is a detrimental to good health

and well being. Obesity is the most prevalent nutritional disorder in prosperous

communities and is the result of an incorrect energy balance leading to an increased

storage of energy, mainly as fat. It is the most common nutritional disorder in western

countries and among the higher income groups in developing countries. The prevalence of

obesity is increasing in most of the populations of world, affecting children, adolescents

and adults (Bar Dayan et al., 2005). Kopelman (2000) suggested that obesity is now so

common within the world’s population that it is beginning to replace under nutrition and

infectious diseases as the most significant contributor to ill health. Obesity is increasing at

an alarming rate throughout the world. Thus, obesity should not be regarded simply as a

cosmetic problem affecting certain individuals, but a serious ailment that threatens global

well being (WHO, 2000).


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Overweight refers to increased body weight in relation to weight, when compared to the

same standard of acceptable or desirable weight. Obesity is defined as an excessively high

amount of fat or adipose tissue in relation to lean body mass (Stunkard and Wadden,

1993). The amount of excess fat, its distribution within the body and the associated health

consequences vary considerably between obese individuals. Obesity may develop at any

age in either sex and as an increasing health problem. Obesity develops over time and once

it develops, is difficult to treat. The excess of fat in men tends to accumulate in the upper

abdomen (Bose, 1995). The site of fat accumulation is considered a predominant factor for

metabolic disorders of obesity (Van Gaal et al., 1988). Several reasons may contribute to

the development of obesity. It is not a single disorder but a heterogeneous group of

conditions with multiple causes.

Recent epidemiological trends in obesity indicate that the primary cause

of the global obesity problem lies in environmental and behavioural changes.

Georges et al. (1991) suggested a larger role for socio-cultural factors in the patterning of

body fat distribution. Mueller and Reid (1979) stated that environmental factors such as

nutrition, stress and exercise have significant effect on subcutaneous fatness. On the other

hand, the role of inherited factors in the origin of obesity is anticipated. Whereas clear

genetic effects exist, these are modified by environmental and behavioral factors

(Pi-Sunyer, 1994). Thus, obesity is multifactorial in origin. In developed countries, the

occurrence of obesity is higher in the lower socio-economic groups, whereas in developing

countries this relationship is reversed (Sobol and Stunkard, 1989).

The worldwide obesity problem can be viewed as a consequence of the substantial

economic, social and cultural problems now observed in developing and newly

industrialized countries. In India the increased levels of obesity is primarily associated

with the transition from rural to urban lifestyle. However, it is evident that this

phenomenon is more profound among the urban populations in comparison to the rural

ones (Venkatramana and Chengal Reddy, 2002). Regardless of its cause, obesity may be

associated with a variety of risks. Obesity causes or exacerbates many health problems,

both independently and in association with other diseases. It is related to the risk for

developing non-insulin dependent diabetes mellitus, hypertension and cardiovascular

disease (Smith et al., 2001). It also creates an enormous psychological burden. Thus,

obesity is associated with a significant increase in morbidity and mortality.


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1.1.1. Classification of Obesity

The fat distribution in the body is identified among the two types of obesity

android and gynoid (Simic et al., 1989).

1.1.1.1. Android

Android is the male type of obesity where excess fat is marked in the upper half of

the body. Android type of obesity is likened to the shape of an apple. The shoulders, face,

arms, neck, chest and upper portion of the abdomen are bloated. The stomach gives a stiff

appearance. The back seems to be erect but the neck is compressed and there will be

protruding chest because of the bulk in the stomach. The lower portion of the body the

hips, thighs and legs are thinner beyond proportion in comparison with the upper part.

In these persons the vital organs affected will be mostly the heart, liver, kidneys and lungs.

Android type of obesity is a major risk for heart damage and heart disease due to high

cholesterol.

1.1.1.2. Gynoid

In this type the lower part of the body has the extra flesh. This type of obesity is

also common to both sexes though females are more affected. Gynoid type of obesity is

similar to the shape of pears. The flesh is somewhat flabby in the abdomen, thighs,

buttocks and legs. The face and neck mostly give a normal appearance. In some persons,

the cheeks may be drawn too. As these persons grow old the whole figure assumes a

stooping posture and the spine is never erect due to the heavy hips and thighs. These vital

organs affected mostly the kidneys, uterus, intestines, bladder and bowls. But the functions

of these organs sometimes have a direct effect on the heart. In this type of obesity,

exercises or dieting will not help appreciably in reducing weight. One should have more

patience and undertake proper treatment to achieve the goal of reducing weight and

preventing further weight again.

1.1.1.3. The third type

Besides android and gynoid, there is one more type of obesity. Some people do not

belong to any of the above types of obesity. Their whole body from head to toe looks like a

barrel. Their gait is more to rolling rather than walking. The fat tissues in their body hinder

the movement of all the internal organs and consequently affect their brisk functioning.

For them any exercise is difficult due to the enormous size of the body. So such person

should follow a strict diet and do plenty of exercise.


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1.1.2. BMI as a Measure for Assessing Obesity

Anthropometrics plays an important role in the assessment of obesity

in conjunction with other sophisticated techniques viz., bioelectrical impedance

analysis (BIA), magnetic resonance imaging (MRI), dual-energy x-ray absorptiometry

(DEXA), isotope dilution, computed tomography (CT), hygrometry and ultrasound

(Van der Kooy and Seidell, 1993). In anthropometry, body mass index (BMI) is the most

commonly used measure of overall obesity (generalized obesity) while circumferences and

skin folds are measures of central obesity (Anne Waldron, 2007). BMI can be considered

to provide the most useful, albeit crude, population level measure of obesity (WHO, 1995;

2000). Body weight is reasonably correlated with body fat, but is also highly correlated

with height. Therefore, weight adjusted for height squared (BMI in kg/m2) is a useful

index to assess overweight and is a fairly reliable surrogate for adiposity. It is calculated

easily from weight and height and it correlates with other measures of body fatness in

children and adolescents. BMI also correlates with markers of secondary complications of

obesity, including current blood pressures, blood lipids and with long-term mortality

(Gidding et al., 1995). According to WHO (2004) classification, BMI<18.5 is under

weight, 18.5 to 24.9 is healthy, 25 to 29.9 is overweight, 30 to 39.9 is obese and >40 is

morbidly obese. Increasing BMI values are correlated with an increased risk of morbidity

and mortality.

1.1.3. Abdominal Obesity

Recently estimates of waist circumference (WC) are gaining increasing importance

as a more useful tool in the assessment of body fat distribution and in the diagnosis of

abdominal obesity (Folsom et al., 1998). Waist circumference (WC) is an indicator of deep

adipose tissue and it is related to fat mass. Waist circumference is a convenient and simple

measurement that is an approximate index of intra-abdominal fat mass and total body fat

(Lean et al., 1996). In addition, waist hip ratio (WHR) is an indicator of the degree of

masculine distribution of adipose tissue. It is now well established that a high WHR

indicates abdominal fat accumulation (Bose and Mascie-Taylor, 1998). Measurements of

impedance (bioelectrical impedance) have recently been introduced and provide accurate

measurements of body fat on most adults. Bioelectrical impedance analysis (BIA) is a safe,

noninvasive, portable method of estimating body composition (Houtkooper et al., 1996).


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Individuals with abdominal obesity are at a great risk for developing diabetes and

atherosclerotic CVD (Ginsberg, 2000; Wang et al., 2005). Abdominal obesity is

commonly associated with hyperinsulinemia, impaired glucose tolerance and

hyperglycemia, as well as increase in plasma triglycerides, small LDL-c particles and

apolipoprotein B and a decrease in HDL-c. Also, abdominal obesity is a major component

of the metabolic syndrome. The diagnosis of abdominal obesity depends on measurement

of WC (Tan et al., 2004).

1.1.4. Factors Associated with BMI and Obesity

1.1.4.1. Demographic Factors: Gender, Age and Ethnicity

a. Gender

Women generally have a higher prevalence of obesity (BMI >30 kg/m2), especially

after the age of 50 years, whereas men usually have a higher prevalence of overweight

(BMI 25 to 29.9 kg/m2) (Flegal et al., 1998; Stam Morega et al., 1999).

b. Age

A BMI increase with age has been documented in several cross-sectional studies

(Boyle et al., 1994; Seidell et al., 1995; Flegal et al., 1998). The older the subjects, the

higher the mean BMI and other prevalence of obesity in both men and women, at least up

to the age of 50 to 60 years (Rolland-Cachera et al., 1991; Seidell et al., 1995).

c. Ethnicity

The prevalence of obesity has been shown to vary across ethnic groups

(Flegal et al., 1998). These differences have been suggested to be partly due to a genetic

predisposition for obesity, which becomes apparent especially when individuals are

exposed to an affluent lifestyle, such as Pima Indians in Arizona or Australian Aboriginals

in an urban environment (WHO, 2000).

1.1.4.2.Socio-cultural Factors: Education and Family Situation

a. Educational Level

The socio-economic gradient in obesity is abundantly accepted in the literature.

Especially in women, a strong inverse association between obesity and socio-economic

status (SES), mostly assessed by educational level, has been reported in numerous affluent

populations (Wamala et al., 1997; Rahkonen et al., 1998; Wardle and Griffith, 2001).


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b. Marital Status

Marital status has been found to be linked with BMI and obesity, although this

association is not well recognized. Several (Khan et al., 1991; Rosmond et al., 1996),

but not all (Tavani et al., 1994) cross-sectional studies have shown married or cohabiting

subjects to have a higher BMI than subjects living alone. Overweight tends to rise after

marriage (Sundquist and Johansson, 1998).

c. Number of Children

Child bearing has been recommended to be a provider to obesity in women, with

pregnancy belonging to the vulnerable period for enlargement of obesity (WHO, 2000).

The effect of child bearing on body weight may be due to environmental factors rather

than being purely biological.

1.1.4.3. Dietary Intake, Physical Activity, Alcohol Consumption and Smoking

It is important to note that weight changes observed in populations over time are

generally so small that they are unlikely to be detected by existing methods for measuring

energy expenditure and energy intake in population (Seidell, 1997; Heitmann, 2000).

a. Food Choices and Dietary Intake

Nutrition is of critical importance in establishing a positive energy balance. Of the

nutritional factors related to obesity, dietary fat intake is widely believed to be the primary

determinant of body fat (Bray and Popkin, 1998). High fat diets have been suggested to

promote obesity by increasing energy intake, further increasing the likelihood of a positive

energy balance and weight gain (Ravussin and Tataranni, 1997; Hill et al., 2000).

b. Physical Activity

Physical activity has been shown to be aversely associated with BMI in numerous

cross-sectional studies (Blokstra et al., 1999) and obese subjects have been observed to be

physically less active than the non-obese (Miller et al., 1990; Cooper et al., 2000).

c. Alcohol Consumption

Epidemiological findings regarding the association of alcohol consumption with

body weight have been controversial. Alcohol is a considerable component of the diet in

many countries, providing about 3 to 9% of daily energy intake (Westerterp et al., 1999).


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Similar to measuring food intake, measuring alcohol consumption is liable to reporting

errors and to being influenced by cultural differences (Caetano, 1998;

De Vries et al., 1999).

d. Smoking Habits

Numerous studies have shown that smoking is associated with lower BMI

(Istvan et al., 1992). In addition, some prospective studies have suggested that smokers

gain more weight than non smokers during the follow up period and also during cessation

of smoking (Coakley et al., 1998).

1.1.5. Complications of Obesity

1.1.5.1. Metabolic-Hormonal Complications

Metabolic syndrome showed some mild adverse reaction, such as type II diabetes,

insulin resistance, hyperinsulinemia, dyslipidemia, hypertension, gout and sleep disorders.

Abnormalities of hormones and other circulating factors employed include cytokines,

ghrelin, growth hormone (GH), hypothalamic-pituitary-adrenal (HPA) axis and leptin and

renin-angiotensin system (Knowler et al., 1990; Sjostrom et al., 1999).

1.1.5.2. Diseases of Organ Systems

The following medical conditions are also more common in obese people than

in those of normal weight: cardiac and vascular disease such as cerebrovascular disease,

congestive heart failure, coronary heart disease, hypertension and thromboembolic disease.

The respiratory system abnormalities are obesity hypoventilation syndrome and sleep

apnea. The reproductive system abnormalities are hormonal complications in females,

males and obstetric complications (Wilson et al., 2002; Delgado-Aros et al., 2005).

1.1.5.3. Cancer

The important consequences (complications) of obesity include increased incidence

of cancer in breast, colon, female reproductive system, cervix, endometrium, ovary, gall

bladder, kidney and prostate (Vainio and Bianchini, 2002; Danaei et al., 2005).

1.1.5.4. Mechanical Complications of Obesity

Arthritis and increased intra-abdominal pressure are a common problem in obese

individuals due to trauma of excess weight bearing (Hart and Spector, 1993;

Cicuttini et al., 1996).


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1.1.5.5. Surgical Complications

Surgical procedures have several risks, particularly if general anesthesia is used.

Morbid obesity increases the risk of post-operative wound infections, increasing the risk

that someone will develop a blood infection or die from overwhelming infection

(Pasulka et al., 1986).

1.1.5.6. Psycho-Social Complications

Multiple aspects of psycho-social functioning include: psychological, social

complications and economic impact (Gortmaker et al., 1996).

1.1.6. Categories of Drugs Used to Treat Obesity

1. Adrenergic agonists

DHEA IIa

: Amphetamine, Methamphetamine.

DEA III : Benzphetamine, Phendimetrazine.

DEA IV : Diethylpropion, Mazindolb, Phentermine.

OTC : Phenylproanolamineb.

2. Serotonin agonists

DEA IV : 1-fenfluramine, d-fentluramine.

Unscheduled : Fluoxelined, Sertraline

d.

3. Combined adrenergic and serotonergic

Agonist : DEA IV : Sibutramine (Merida).

4. Drugs effecting absorption

Unscheduled : Orlistat and Acarbosed.

5. Other off-label and over the counter drugsd

Prescription drugs (off-label): Bromocriptine, Bupropion, Diazoxide, Metformin,

Topiramate, Zonisamide and Felbamate.

OTC (Over the counter) drugs: Chromium picolimate, Cimetidine,

Dihydro-epiandrosterone (DHEA), Ephedrine-caffeine, Gamma-amine butyric acid

(GABA) and Nicotine.


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a DEA schedule II drugs are not recommended for obesity treatment.

b Not available in the United States.

c Withdrawn from the market by the manufacturer, September, 1997.

d Not approved for obesity treatment.

6. Diet supplements

Diet supplements are choline-inositol, cayenne, cinnamon, ephedra-coffee bean

extracts, Garcinia cambogia, ginger, green tea, guarana, mustard seed and Spirulina.

1.1.7. Mechanism of Action of Anti-obesity Drugs

Any treatment for controlling obesity will decrease the cost of health services.

Obesity could affect anesthesia procedure where high concentrations of anaesthetic drug

accumulate in adipose tissue. Postoperative complication may prevent some operations in

obese patients such as hip replacement and aneurysm repair (MacConnachie, 1999).

There are two main categories of anti-obesity drugs: anti-obesity drugs that act on

the gastrointestinal system (pancreatic lipase inhibitors) and central nervous system mainly

to suppress appetite. The first kind of anti-obesity drug inhibits pancreatic and gastric

lipase. It decreases ingested triglyceride hydrolysis that produces a dose-dependent

reduction in dietary fat absorption which in turn leads to weight loss. It is a peripheral

mode of action, is including weight loss by selectively inhibiting gastrointestinal lipase

activity, thereby reducing the absorption of dietary fat (McMahon et al., 2000;

Wirth and Krause, 2001). The second kind of anti-obesity drug promotes a sense of satiety

through its central action as a serotonin, dopamine and norepinephrine reuptake inhibitor,

which induces weight loss by enhancing satiety and increasing metabolic rate. It may also

alleviate against the fall in thermogenesis through stimulation of peripheral norepinephrine

receptors. It is, in simple terms, an anorectic or appetite suppressant, that reduces the

desire to eat (Van Gall et al., 1998; Gokcel et al., 2001; Serrano-Rios et al., 2002).

The recently developed anti-obesity drug (Rimonabant) also acts centrally on the brain and

decreases appetite. It may also act peripherally by increasing thermogenesis and therefore

increasing energy expenditure (Lowell and Spiegelman, 2000; Hainer, 2011).


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1.1.8. Contraindications to the Use of Anti-obesity Drugs

Centrally acting drugs are not recommended for patients who are concurrently

taking other selective serotonin reuptake inhibitors. It is therefore crucial to confirm that

such drugs have not already been prescribed. It is probably also unwise to co-prescribe

with tricyclic anti-depressants, monoamine oxidase inhibitors and lithium, all of

which may potentiate the central effects of serotonin with adverse results. Combination

therapy with anti-obesity drugs is contraindicated because of the absence of evidence for

synergy between the two drugs and lack of information about safety (Harvey et al., 1999;

Members of Nutrition Committee, 2003).

1.1.9. Current Criteria for the Evaluation of New Anti-obesity Drugs

Before a new anti-obesity compound can be approved for use it must be proven to

be both safe and effective. The American food and drugs administration (FDA) and the

European agency for the evaluation of medicinal products (EMEA) set the guidelines by

which drugs currently under development are assessed (Halpern and Mancini, 2003).

Once an anti-obesity drug has passed through initial trials to test its safety and establish

effective and tolerable doses, the drug’s clinical efficacy must be tested against a placebo

in larger scale, longer term randomized double blind trials.

Both the FDA and EMEA understandably demand that any anti-obesity drug

should produce significantly greater weight loss compared to placebo control over any

trial. The secondary outcome of anti-obesity drug trials is to ensure that this weight loss in

sustained and that it produces a significant reduction in risk factors for a number of obesity

related co-morbidities (e.g., fasting blood glucose, HbA1c, insulin, total plasma cholesterol,

LDL-cholesterol, triglycerides, uric acid and blood pressure). The FDA also demands that

drugs reduce total body fat mass and alter body fat distribution. Finally, drug induced

weight loss should have made a positive impact on life style (Halford, 2004).

1.1.10. Future Treatment of Obesity

The maximal weight loss achievable with any current dietary or pharmacological

strategy for treating obesity varies with the individual but appears to be no more than 10%

of initial weight. As this threshold is approached, or perhaps as the time spent below initial

weight increases, physiological mechanisms acting to preserve body fat mass cause a


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progressive increase in appetite and decrease in energy expenditure. These regulatory

responses prevent further weight loss and make maintenance of achieved weight loss

difficult. It is now appreciated that the long term regulation of adiposity involves both

peripheral signals that relay information about adipose tissue mass to the CNS and

opposing circuits in the hypothalamus that regulate appetite and energy expenditure

(Schwartz et al., 2000; Langroudia et al., 2011). To improve the pharmacological options

for treating obesity, it will be necessary to intervene at key points within this regulatory

network.

1.2. LITERATURE PERTINENT TO GERIFORTE

Ancient medical literature on ayurveda gives a vivid and comprehensive

description of this group of plant medicines which are tonic, prevent disease and

postpone ageing. The indigenous drug formulation in tablet form, geriforte

(M/s. Himalaya Drug Co., Bangalore, India), is a comprehensive compound of rasayana

drugs, the principal component of which is chyavanprash. It also contains such nervine

tonics as brahmi, ashwagandha, herbal and mineral extracts known for their metabolic and

tonic properties and is prepared in the juices and decoctions of various reconstituents,

hepatic stimulants and digestives (Kishore et al., 1983; Upadhyaya et al., 1990). Thus,

geriforte, an indigenous herbo-mineral compound, is a mixture of several major and minor

herbal extracts and minerals. The major constituents are discussed below and minor

constituents are detailed in appendix – I and II.

Geriforte induces cellular regeneration (Lobo et al., 1975), increases hormonal

utilization (Kishore et al., 1983), regulates enzymes and bio-amine metabolism

(Bardhan et al., 1985; Upadhyaya et al., 1988), inhibits the formation of lipofuscin in

ageing rats (Sharma et al., 1991; 1992) and maintains the cytoprotective and

immunomodulating activities (Bansal et al., 2001). The clinical significance of this remedy

has already been reported in the management of anxiety and psychosomatic disorders

(Singh et al., 1978; Dubey et al., 1984), maintains positive nitrogen balance and cures

menopausal symptoms (Bardhan et al., 1985). It is also reported to act on the

cardiovascular, sexual and other systems and helps to correct the metabolism of proteins,

fat, carbohydrates, serum cholesterol, triglycerides, phospholipids, etc. (Singhal, 1978).


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1.2.1. Literature Pertinent to Chyavanprash

Chyavanprash is an ancient ayurvedic health tonic, widely used in India, as a

rejuvenative, energizer and immunity booster. It is often called the elixir of life due to its

numerous nutritional properties and benefits to the body (www.himalayahealthcare.com).

1.2.1.1. Composition

Chyavanprash is a brown coloured, sticky paste with the consistency of jam and a

sweet/sour/spicy taste. It includes the ingredients: fresh amla fruit, sugar, honey, ghee,

long pepper, sesame oil, giant potato, cardamom, bamboo manna, Indian kudzu, winter

cherry, Asparagus, cinnamon bark, dashmool, country mallow, wild green gram, wild

black gram, galls, feather foll plant, raisins, Ceylon-cow plant, Irish root,

Chebulic myrobalan, round zedoary, nut grass, spreading hogweed, blue Egyptian water

lily, Malabar nut, liquorice, tiger's claw or ice plant, sandal wood, clove, Chinese

cinnamon, cobra's saffron and potassium sorbate (www.himalayahealthcare.com).

1.2.1.2. Benefits

Regular intake of chyavanprash strengthens digestion, absorption and assimilation

of food. It is also beneficial to the heart and the brain cells. It is considered a memory

booster. It also works as an antioxidant, thus slowing down the aging process. It is

believed that chyavanprash purifies blood, eliminates toxins and is beneficial to liver.

It also improves muscle tone by enhancing protein synthesis. It is especially beneficial for

cough and asthma patients. It enhances fertility and keeps menstruation regular

(www.himalayahealthcare.com).

1.2.2. Literature Pertinent to Capparis spinosa

Family : Capparidaceae

English Name : Caper

Sanskrit Name : Karira

Hindi Name : Kachra, Kabra, Karer

Tamil Name : Kariyal

1.2.2.1. Habitat

It grows in Afghanistan, West Asia, Europe, North Africa and Australia. In India it

is grown from Punjab and Rajasthan upto the Deccan Peninsula (Nadkarni and Nadkarni,

1976).


http://en.wikipedia.org/w/index.php?title=Malabar_nut&action=edit&redlink=1

http://en.wikipedia.org/wiki/Digestion

http://en.wikipedia.org/wiki/Absorption

http://en.wikipedia.org/wiki/Assimilation_(biology)

http://en.wikipedia.org/wiki/Blood

http://en.wikipedia.org/wiki/Liver

http://en.wikipedia.org/wiki/Cough

http://en.wikipedia.org/wiki/Asthma

http://en.wikipedia.org/wiki/Fertility

http://en.wikipedia.org/wiki/Menstruation

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1.2.2.2. Morphology Description

C. spinosa is a prostrate, glabroscent, polymorph shrub or climber armed with

divaricate light yellow thorns, occurring in dry rocky and stony soils. Branches are terete

glabrous or pubescent. Leaves are variable in texture, orbicular to elliptic, base rounded

and apex mucronate. Flowers are white, solitary and auxiliary. The berry is ellipsoid,

ovioid or obovoid and pericarp. The seeds are 3 to 4 mm in diameter, globose, smooth and

brown (Lange et al., 1982).

1.2.2.3. Principal Constituents

The cortex and leave contains stachydrine and 3-hydroxystachydrine. The root

contains glucobrassicin and neoglucobrassicinand 4-methoxy-glucobrassicin. The crude

extract of the flower bud contains isothiocyanates, thiocyanates and sulphides

(Schraudolf, 1989; Brevard et al., 1992).

1.2.2.4. Pharmacology

C. spinosa was reported to have a number of potentially useful medicinal attributes

including anti-oxidative (Germano et al., 2002), anti-fungal (Ali-Shtayeh and

Abu-Ghdeib, 1999), anti-hepatotoxic (Gadgoli and Mishra, 1999), anti-inflammatory

(Rossi et al., 1988), anti-diabetic (Yaniv et al., 1987) and anti-obesity

(Lemhadri et al., 2007). In Morocco, this plant was traditionally used in diabetes control

and treatment (Eddouks et al., 2002) and has hypoglycemic activity (Eddouks et al., 2004;

Eddouks et al., 2005; Hashemnia et al., 2012).

1.2.2.5. Indications

The plant is also credited with anti-tubercular property. The bark is bitter, used as a

diuretic and an expectorant. It is given for spleen, renal and hepatic complaints. The plant

extract is administered to treat senile pruitis, itching and other ailments associated with old

age and anxiety neurosis (Gadgoli and Mishra, 1999; Upadhyay, 2011).

1.2.3. Literature Pertinent to Cichorium intybus

Family : Asteraceae

English Name : Chicory, Succory, Wild endive

Sanskrit Name : Kasani, Hinduba, Kasni

Tamil Name : Kaasini


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1.2.3.1. Habitat

It is native to the temperate parts of the old world and is found in Punjab and

Andhra Pradesh. It is cultivated in Bihar, Punjab, Himachal Pradesh, Assam, Maharashtra,

Gujarat, Tamil Nadu, Orissa, Andhra Pradesh and Kerala (Bruneton, 1999).


C. intybus is an erect perennial herb, with a fleshy tap root; the leaves are broadly

oblong, oblanceolate or lanceolate, crowded at the base and arranged spirally on the stem;

the flowers are blue fading to white; the fruits are achenes which are smooth, 5 angled,

pale brown to black and crowned with a ring of pappus scales (Bruneton, 1999).


The active compounds in C. intybus are inulin, sesquiterpene lactones, vitamins,

minerals, fat, mannitol and latex. Fructans have been extracted from chicory roots

(Finke et al., 2002). Chemical constituents of C. intybus are identified as α-amyrin,

taraxerone, baurenyl acetate and β-sitosterol (Du et al., 1998).


C. intybus inhibits mast cell-mediated immediate type allergic reactions in vivo and

in vitro (Kim et al., 1999). Inulin decreases serum triglycerides by decreasing fatty acid

synthesis and reducing production of low density lipoproteins (LDL) (Williams, 1999).

C. intybus has been found to inhibit prostaglandin E-2 and cyclooxygenase-2 (COX-2)

(Cavin et al., 2005). C. intybus derived β-fructans have been shown to exert cancer

protective effects in animal models (Hughes and Rowland, 2001; Pool-Zobel, 2005).

Oligofructose, inulin and selective fermentable chicory fructans have been shown to

stimulate the growth of bifido bacteria, which are regarded as beneficial strains in the

colon (Reddy, 1999; Chow, 2002).


The seeds are reported to be carminative and cordial and used as a brain tonic.

They are useful in headache, asthma and for checking bilious vomiting

(Ahmed et al., 2003).


Introduction

16 | P a g e

1.2.4. Literature Pertinent to Berberis aristata

Family : Berberisdaceae

English Name : Indian berberry, Tree turmeric

Sanskrit Name : Daruharidra, Darvi, Darurajani

Hindi Name : Darhald

Tamil Name : Mullukala, Usikkala

1.2.4.1. Habitat

Berberis aristata is a spinous shrub native to mountainous parts of North India and

Nepal. These shrubs are distributed throughout the Himalayas, from Bhutan to Kunawar

(altitude 6 to 10,000 ft), Nilgiri hills (altitude 6 to7,000 ft) and Sri Lanka (altitude 6 to

7,000 ft) (Kirtikar and Basu, 1995).


B. aristata is an erect, glabrous, spinescent shrub with obovate to elliptic, sub-acute

to obtuse and entire or toothed leaves. The flowers are yellow and in corymbose racemes.

The fruits are oblong-ovoid or ovoid and bright red berries (Kirtikar and Basu, 1995).

1.2.4.3. Principal constituents

The alkaloids in the bark and root bark of Berberis aristata are berberine,

berbamine, karachine, palmatine, oxyacanthine and oxyberberine (Rahman and

Ansari, 1983). The fruit contains 0.64 per cent tannin and 0.37 per cent pectin.

Its vitamin C content is only 4.60 mg per 100 ml of juice (Parmar and Kaushal, 1982).


B. aristata has been traditionally used in all types of inflammations, throat

infections, wound healing, dysentery, uterine and vaginal disorders. It is well known for its

anti-inflammatory and immunopotentiating property. Berbamine effectively inhibits

chemically-induced hepatocarcinogenesis. Preliminary reports indicate that it possesses

anti-cancer activity as tested against mouse leukemic L1210 cells, human hepatoma cells

and colon cancer cells. It is postulated that its anti-cancer activity may be due to its

COX-II inhibitory property (Fukuda, 1999). The other uses of Berberis aristata are as

cooling laxative diaphoretic and useful in rheumatism. The dried extract of the roots are

applied externally to the eyelids to cure ophthalmia and other eye diseases

(Jain and Singh, 1994).


Introduction

17 | P a g e


The root possesses anti-bacterial and anti-inflammatory activities. The root bark is

very useful in periodic neuralgia, menorrhagia (Dutta and Panse, 1962) and has effective

anti-diabetic/hypoglycemic chemical principle(s) which possess insulin triggering and

insulin like activities (Akhtar et al., 2008).

1.2.5. Literature Pertinent to Adhatoda vasica

Family : Acanthaceae

English Name : Malabar nut

Sanskrit Name : Shwetavasa, Vasa, Vasaka

Hindi Name : Arusa, Baansa, Adulsa

Tamil Name : Adatodai

1.2.5.1. Habitat

The plant is distributed throughout India upto an altitude of 1,300 m and mainly

found in sub-Himalayan regions; also found in Nepal, Pakistan, Myanmar and Germany

(Kapoor, 2001).


A. vasica is a small evergreen and sub-herbaceous bush. The leaves are 10 to 16 cm

in length, minutely pubescent and broadly lanceolate. The inflorescence is dense, short

pedunculate, bractate and spike terminal. The fruit is 4 seeded small capsules. The stomata

in the plant are elongated and oval in shape (Kapoor, 2001).


The chief alkaloid present in the leaves of A. vasica is a quinazoline alkaloid and

vasicine. Vasicine is accompanied by α-vasicinone, deoxyvasicine and maiontone.

The roots of the plant contain vasicinolone, vasicol, peganine, hydroxy oxychalcone and

glucosyl oxychalcone. The flowers of the plant contain β-sitosterol-D-glucoside,

kaempferol, glycosides of kaempferol and queretin (Kapoor, 2001; Sharma et al., 2012).


A. vasica is commonly used for treatment of respiratory complaints. The leaves are

boiled and taken orally for fevers (Sane et al., 1995); warmed leaves are applied externally


Introduction

18 | P a g e

for rheumatic pains and dislocation of joints. The powder boiled in seasame oil is used for

ear infection and to stop bleeding, dried leaves powder is used for stomach acidity,

decoction of leaves are used to facilitate childbirth or induce abortion (Shah et al., 1999).

A paste of leaves is applied to the abdomen to treat urinary disorders

(Claeson et al., 2000). The leaves and flowers used as vegetables, an expectorant and

febrifuge (Pandey and Chaturvedi, 1969).


In ayurveda, mostly the leaves are used in the treatment of respiratory disorders.

The alkaloids, vasicine and vasicinone present in the leaves, possess respiratory stimulant

activity. Low concentrations of vasicine induce bronchodilation and relaxation of the

tracheal muscle. Higher concentrations of vasicine offer significant protection against

histamine-induced bronchospasm in guinea pigs (Gupta et al., 1977).

1.2.6. Literature Pertinent to Withania somnifera

Family : Solanaceae

English Name : Winter cherry

Sanskrit Name : Ashvagandha, Hayahvaya, Vajigandha

Hindi Name : Asgandh

Tamil Name : Amukkara

1.2.6.1. Habitat

It is tomentose under shrub. The leaves are ovate, sub-acute and pubescent.

Flowers are greenish or lurid yellow. Berry is red enclosed in the inflated calyx. The seed

are reniform and yellow (Owais et al., 2005).


W. somnifera is an erect, evergreen and tomentose shrub. The roots are stout, fleshy

and whitish brown; the leaves are simple ovate and glabrous; the flowers are

inconspicuous and greenish or lurid yellow; the berries are globose, orange-red when

mature, enclosed in the persistent calyx and reniform seeds (Owais et al., 2005).


The roots of W. somnifera contain several alkaloids, glycosides, a few flavonoids

and reducing sugars (Ganzera et al., 2003) and are also rich in iron (Mishra et al., 2000).


Introduction

19 | P a g e

Biochemically heterogeneous alkaloids are present including cuscohygrine, anahygrine,

tropine, pseudotropine and anaferine. The plant has steroidal lactones-withanolides and

withaferin, which are estrogenic compounds (Dhuley, 2000).


The chemical composition, pharmacological and therapeutic efficacy of this plant

has been well established (Dhuley, 2000). Different investigators reported that

W. somnifera possess anti-serotogenic, hyperlipidemia, Parkinson’s disease

(Gupta and Rana, 2007), anti-cancer, anabolic activity and beneficial effects in the

treatment of arthritis, geriatric problems (Asthana and Raina, 1989) and stress

(Grandhi et al., 1994). Further the plant has been reported to have anti-inflammatory,

anti-tumour, anti-stress, antioxidant, immunomodulatory, haematopoietic and rejuvenating

properties (Gautam et al., 2004; Rasool and Varalakshmi, 2006). It is one of the most

commonly used herbs, not only as an anti-stress and adaptogenic agent, but is also known

to increase life span and delay ageing (Bhatnagar et al., 2005).


W. somnifera is used in asthma and as a uterine sedative. The total alkaloids

showed relaxant and anti-spasmodic effects against several spasmogens on

intestinal, uterine, bronchial, tracheal and blood vascular muscles (Dhuley, 2000;

Harikrishnan et al., 2008).

1.2.7. Literature Pertinent to Asparagus recemosus

Family : Liliaceae

English Name : Asparagus

Sanskrit Name : Shatavari

Hindi Name : Satavar, Satmuli

Tamil Name : Tannirvittan, Ammalkodi, Kadumulla, Niliechedi

1.2.7.1. Habitat

Asparagus recemosus is found throughout tropical Africa, Java, Australia, India,

Sri Lanka and southern parts of China. In India it is found in plains to 4,000 ft height

(Anonymous, 1987).


Introduction

20 | P a g e


It is an under-shrub, climbs up to 1 to 3 m height, with stout and creeping root

stock. The root occurs in clusters or fascicle at the base of the stem with succulent and

tuberous rootlets. The stem is scandent, woody, triquetros, striate, terete and climbing.

The spines are long, sub-recurved or straight. The flowers are solitary or fascicles and

simple or branched racemes of 3 cm long. The berry is globular or obscurely 3 lobbed and

purple-reddish, seeds hard with brittle testa (Anonymous, 1987).


The major active constituents of root extract A. racemosus are steroidal saponins

namely shatavarins (Hayes et al., 2006) apart from alkaloids, flavonoids, sterols and

terpenes (Bopana and Saxena, 2007). Other active compounds such as quercetin, rutin and

hyperoside are found in the flowers and fruits; while diosgenin and quercetin-3

glucuronide are present in the leaves (Anonymous, 1987).


A. racemosus has been used in ayurveda as a galactagogue, aphrodisiac, anodyne,

diuretic and anti-spasmodic (Sharma et al., 2000). Both aerial parts and roots have amylase

and lipase activities (Dange et al., 1969). Aerial parts have anti-cancer activity in human

epidermal carcinoma of the nasopharynx (Dhar et al., 1968). Further, several formulations

containing A. racemosus have been reported to possess adaptogenic activity

(Bhattacharya and Chakrabarti, 2004). The methanolic extract of fresh roots of

A. racemosus showed significant protection against cold restraint stress induced gastric

ulcers (Sairam et al., 2003).


The roots have oleaginous, cooling, anti-spasmodic, indigestible, appetizer,

alliterative, asphrodisiac, galactagogue, astringent, anti-diarrhoeatic, anti-dysenteric,

laxative properties and is useful in tumours, inflammations, diseases of blood,

throat complaints, tuberculosis, leprosy and kidney troubles (Mandal, 1998;

Venkatesan et al., 2005; Raghav and Kasera, 2012).


Introduction

21 | P a g e

1.2.8. Literature Pertinent to Glycyrrizha glabra

Family : Fabaceae

English Name : Liquorice, Licorice

Sanskrit Name : Yashti-madhuh, Yashti-madhuka

Hindi Name : Jothi-madh, Mulhatti

Tamil Name : Adhimadhuram

1.2.8.1. Habitat

It grows in the sub-tropical and warm temperate regions of the world, chiefly

in Mediterranean countries and China in rich soil to a height of 4 to 5 ft

(Olukoga and Donaldson, 1998).


Glycyrrizha glabra has oval leaflets, white to purplish flower clusters and flat pods.

Below ground, the G. glabra plant has an extensive root system with a main taproot and

numerous runners. The main taproot, which is harvested for medicinal use, is soft, fibrous

and has a bright yellow interior (Olukoga and Donaldson, 1998).


The major bioactive constituent of rhizome is a triterpenoids saponin, glycyrrhizin,

glycyrrhizinic acid, glabrin A and B, glycyrrhetol, glabrolide, isoglabrolide,

isoflavones, coumarins, triterpene sterols, etc. (Vaya et al., 1997; Obolentseva et al., 1999;

Tamir et al., 2001; Zhang and Ye, 2009).


Glycyrrhizin, a glycoside obtained from G. glabra was studied for its anti-arthritic

and anti-inflammatory effect on formaldehyde induced rat-paw edema in adrenalectomised

rats. It was found to potentiate the anti-arthritic action of hydrocortisone in rats. It has been

traditionally used for respiratory, gastrointestinal, cardiovascular, genitourinary, eye and

skin disorders and for its anti-viral effects (Fiore et al., 2005). The anti-ulcerogenic

action of G. glabra and its consumption as a food ingredient has also been reported

(Isbrucker and Burdock, 2006).


Introduction

22 | P a g e


G. glabra is used for the treatment of asthma, acute and chronic bronchitis and

chronic cough. It is a mild anti-inflammatory for arthritis and rheumatism and is used to

treat gastric, duodenal and esophageal ulceration (Das et al., 1989; Krausse et al., 2004).

1.2.9. Literature Pertinent to Centella asiatica

Family : Apiaceae

English Name : Indian pennywort, Centella, Gotu kola

Sanskrit Name : Mandukaparni, Brahmi, Mandukig, Divya

Hindi Name : Brahma-manduki, Khulakhudi, Mandookaparni

Tamil Name : Vallaarai

1.2.9.1. Habitat

Centella asiatica is indigenous to South-East Asia, Madagascar, South Africa,

South-East U.S., Mexico, Venezuela, Columbia and Eastern South America. It is found in

abundance in moist, sandy or clayey soils and other waste places throughout India upto an

altitude of 600 m (Anonymous, 1992).


C. asiatica is a slender, tender and faintly aromatic herb. Stems are prostrate, often

reddish, striated and rooting at the nodes. The leaves are 1 to 3 from each node of the

stems. Flowers are in fascicled umbels, each umbel consisting of 3 to 4 white to purple or

pink and sessile flowers. Fruits are 4 mm long, oval to globular in shape and hard with

thickened pericarp (Kirtikar and Basu, 1987).


C. asiatica contains several active constituents, of which the most important are the

triterpenoid saponins, including asiaticoside, centelloside, madecassoside and asiatic acid.

In addition, Centella contains other components, including volatile oils, flavonoids,

tannins, phytosterols, amino acids and sugars (Rastogi and Mehrotra, 1993;

Schaneberg et al., 2003).


C. asiatica has been used in traditional Indian medicine for various pathological

disorders and in particular for the healing of wounds and for leprosis (Bonte et al., 1994).


Introduction

23 | P a g e

In the ayurvedic system of medicine, it is also recommended in chronic diseases and as a

brain tonic in various mental disorders. It is listed officially in the Chinese pharmacopoeia

and used as an anti-pyretic, diuretic and in the treatment of icterus, diarrhea, ulcerations,

eczema and traumatic diseases (Tang and Eisenbrand, 1992).


The plant is used as an anti-dote to cholera. Internally it has been valued as a tonic

and is used in bronchitis, asthma, leucorrhoea, kidney troubles and dropsy.

A decoction of very young shoots is given for haemorrhoids (Allegra et al., 1981;

Montecchio et al., 1991).

1.2.10. Literature Pertinent to Shilajeet

English Name : Jew’s pitch, Vegetable asphalt

Sanskrit Name : Shilajit, Silajit, Silaras

Hindi Name : Ral-yahudi, Silajita

Tamil Name : Perangyum, Uerangyum

1.2.10.1. Meaning and Source

In Sanskrit, shilajit means winner of rock. Another meaning is sweat of the rock

(Mukherjee, 1992). The charaka samhita states that, stones of metal like gold etc., in the

mountains get heated up by the sun and the exudates that come out of them in the form of

smooth and clean gum is called Cilajatu. Shilajit is a blackish-brown exudation, of variable

consistency, obtained from steep rocks of different formations found in the Himalayas at

altitudes from 1,000 to 5,000 m, from Arunachal Pradesh in the east to Kashmir in the

west. It is also found in Afghanistan, Nepal, Bhutan, Pakistan, China, Tibet and USSR

(Jaiswal and Bhattacharya, 1992).

1.2.10.2. Chemistry

The general appearance of shilajit is that of a compact mass of vegetable organic

matter composed of a dark-red gummy matrix interspersed with vegetable fibers, sand and

earthy matter. Chemical analysis shows that it contains besides gums, albuminoids, traces

of resin and fatty acid; a large quantity of benzoic and hippuric acids and their salts

(Chopra et al., 1958).


Introduction

24 | P a g e


It gives more energy, relieve digestive problems, increase sex drive, improve to

memory etc. It is an effective remedy in arthritic conditions like rheumatoid arthritis,

osteoarthritis, gout, back pain and works as anti-inflammatory substance

(David and Vincent, 2001). It is also used in disorders like depression, mental stress,

epilepsy and mental fatigue (Dash, 1991; David and Vincent, 2001). It helps liver to work

normally helping in proper secretion of all the juices and enzymes important for proper

metabolism in body (Chopra et al., 1958). It is used to maintain physical and mental

strength and to maintain youth and to attain long life (David and Vincent, 2001;

Lad, 2002).


Herbal actions are alterative, diuretic, lithotroptic, antiseptic, tonic, rejuvenative

(David and Vincent, 2001). Other actions include anodyne, anti-helminthic and blood

sugar reducer (Halpern, 2003). It also has a laxative effect and has absorbing and purifying

properties (Ghosal et al., 1991).

1.2.11. Literature Pertinent to Terminalia chebula

Family : Combretaceae

English Name : Chebulic myrobalan, Ink nut

Sanskrit Name : Haritaki, Abhaya, Pathya

Hindi Name : Harad

Tamil Name : Kadukkai

1.2.11.1. Habitat

It is found throughout the greater parts of India (Sharma and Dash, 1998).


T. Chebula is a tree with a rounded crown and spreading branches. The bark is

dark-brown, often longitudinally cracked; the leaves are ovate or elliptic with a pair of

large glands at the top of the petiole; the flowers are yellowish white, in terminal spikes;

the drupes are ellipsoidal, obovoid or ovoid and yellow to orange-brown; the seeds are

hard and pale yellow (Sharma and Dash, 1998).


Introduction

25 | P a g e


The most important part of the plant used, is the fruit, either fresh or dried

T. chebula contains many polyphenolic compounds including chebulagic, chebulinic acid,

gallic acid (GA), ellagic acid (EA) and corilagin (CG) (Worasuttayangkurn, 2001).


T. Chebula is useful in treating constipation and is a gentle bowel cleaner. It helps

to maintain regular bowel movement and ensures the comfort of smooth evacuation. The

fruits are used in heart diseases, respiratory diseases, pain, fever, inflammation, piles,

anemia, diabetes and gastric disorders (Chatterjee and Pakrasi, 2000; Suchalatha and

Shyamala Devi, 2005). T. chebula has been reported to exhibit a variety of biological

activities, such as anti-diabetic (Sabu and Kuttan, 2002; Senthilkumar et al., 2006;

Chattopadhyay and Bhattacharyya, 2007), anti-cancer (Saleem et al., 2002; Chattopadhyay

and Bhattacharyya, 2007), anti-mutagenic (Kaur et al., 2002) and anti-viral activity

(Ahn et al., 2002; Chattopadhyay and Bhattacharyya, 2007).


T. chebula has been studied for its antioxidant (Chen et al., 2003), anti-microbial

(Burapadaja and Bunchoo, 1995) and anti-cancer (Saleem et al., 2002) activities.

Recently, it was reported that oral administration of the extracts from T. chebula reduced

the blood glucose level in normal and in alloxan-diabetic rats (Sabu and Kuttan, 2002).

1.2.12. Literature Pertinent to Makardhwaj

English Name : Sulphide of mercury

Sanskrit Name : Makardhwaj

HgS is one of the most useful mercury salts. Its red form is crystalline and black

form is amorphous (Lee, 1996). The red form is used as colouring material. In Indian

ayurvedic medicine makardhwaj, a combination of HgS, Au and Au2S has been known to

be in use for long in the treatment of a number of diseases. Makardhwaj is a well known

inorganic preparation of the ayurvedic pharmacopoeia. Chemically, it is red sulphide of

mercury and gold in uncombined form. It is a sublimed product made from pure mercury,

sulphur and gold (www.himalayahealthcare.com).


Introduction

26 | P a g e

In the majority of cases, it is mixed with various drugs called anupana or adjuvants.

In cases of indigestion and diarrhea, makardhwaj is mixed with powdered bael fruit

(Aegle marmelos); in cases of fever and cough it is given with the juices of ginger, betel

(Piper betel) and tulsi leaves (Ocimum tenuiflorum). Generally, honey is used in the

absence of proper adjuvants. The medicine can be used both for adults and children, the

dosage being regulated according to age. Makardhwaj, when taken regularly, is believed in

indigenous systems of medicine, to be a wonderful tonic and is said to increase longevity

in a patient (www.himalayahealthcare.com).

A valuable tonic for debilitating conditions and convalescent patients after acute

illness, in failing circulation and cardiac asthenia; it increases the red blood corpuscles and

improves general nutritional status. It is also used as a laxative with good results

particularly in those cases when there are visceroptosis and a tonic condition of the

gastrointestinal tract (www.himalayahealthcare.com).

1.2.13. Literature Pertinent to Asparagus adscendens

Family : Asparagaceae

English Name : Asparagus

Sanskrit Name : Shweta musli

Hindi Name : Safed musli, Khairuva

Tamil Name : Tilapane, Taltad

1.2.13.1. Habitat

It grows in the Western Himalayas, Himachal Pradesh and Kumaun upto an

altitude of 1,500 m (Rawat et al., 2010).


It is a sub-erect and prickly shrub with white tuberous roots. Stem is sub-erect,

terete and smooth. Cladodes are dense, slender, filiform, terete, soft and sub-erect or

curved. Racemes are many flowered. The berry is one seeded (Rawat et al., 2010).


Steroidal glycosides and several compounds like 3-heptadecanone,

8-hexadecenonoic acid, methyl pentacosanoate, palmitic acid and stearic acid were

identified in this plant (Rawat et al., 2010).


Introduction

27 | P a g e


All parts such as stem, seeds and rhizomes of this species are very important in

Indian traditional medicinal remedies for treatment of spermatorrhoea, chronic

leucorrhoea, dysentery, asthma and fatigue. The rhizome powder is given as a nutritive

tonic with milk in case of seminal weakness and impotence (Kapoor, 2001). The rhizome

extracts contain a large number of very important steroids, glycosides, saponins and

essential oil (Tandon and Shukla, 1995; Dinan et al., 2001). It has also been identified as

one of the drug to control the symptoms of AIDS (Trivedi and Upadhyay, 1993).


The tuberous root are used as tonic and said to be useful in general debility

(Rawat et al., 2010).

1.2.14. Literature Pertinent to Myristica fragrans

Family : Myristicaceae

English Name : Nutmeg, Mace

Sanskrit Name : Jatiphala, Jatiphalam, Jatikosha, Jatipartri, Jatipatra

Hindi Name : Jaiphal, Jayapatri, Javitri

Tamil Name : Jadikkai

1.2.14.1. Habitat

The plant is a native of Moluccas, now cultivated in many tropical countries of

both hemispheres. In India, it is grown in Tamil Nadu, Kerala and Assam (Datla, 1988).


M. fragrans is a dioecious or occasionally monoecious evergreen and aromatic tree.

The bark is grayish black. The leaves are elliptic or oblong-lanceolate and coriaceous.

The flowers are in umbellate cymes, creamy yellow and fragrant. The fruits are broadly

pyriform or globose. The seed are broadly ovoid, albuminous, with a shell-like purplish

brown testa and covered by a red, fleshy aril (Tiwari et al., 1990).


M. fragrans contains a volatile oil, starch (Santos et al., 1997; Olajide et al., 1999)

and amylodextrin (Datla, 1988).


Introduction

28 | P a g e


Nutmeg is popular as a spice and also possesses various therapeutic properties.

M. fragrans is used for both culinary and medicinal purposes. M. fragrans is used for

treating diarrhea, mouth sores and insomnia (Van and Cox, 1994). Compounds isolated

from the seeds of this plant have been reported to possess strong platelet anti-aggregatory

activity (Venton et al., 1991; Janssens et al., 1990). M. fragrans contains essential oils,

many of which have been reported to possess analgesic and anti-inflammatory properties

(Santos et al, 1997; Olajide et al., 1999). Also, M. fragrans prevents hypercholesterolemia

and atherosclerosis (Sharma et al., 1995). The ethanolic and petroleum ether extract of the

nutmeg significantly produced hypolipidemia, hypoglycemic and anti-diabetic activity

(Ram et al., 1996; Somani and Singhai, 2008).


M. fragrans is reported to be an expectorant, vermifuge, aphrodisiac and as a

nervine used by psychiatrists (Kalbhen, 1971). M. fragrans is used in folk medicine for the

treatment of rheumatism. It has anti-inflammatory, anti-fungal and anti-bacterial properties

(Ozaki et al., 1989).

1.2.15. Literature Pertinent to Eclipta alba

Family : Asteraceae

English Name : Kadimulbirt

Sanskrit Name : Bhringraj

Hindi Name : Bhagra

Tamil Name : Karasalankanni

1.2.15.1. Habitat

Eclipta alba grows as a common weed throughout India, ascending upto 6,000 ft

on the hills (Chopra et al., 1996).


It is a small shrub that is found growing horizontal on the ground. The stem is of

black color. Leaves are 1 to 4 inch long and 0.5 to 1 inch broad. It is ovate in shape and the

edges of leaves are toothed. Petiole is small from apex of which arises a white colored

flower. Fruit is of 0.1 inch in length having hairs on the posterior surface. Seeds are small

and very similar to that of cumin seeds (Chopra et al., 1996).


Introduction

29 | P a g e


The herb is a rich source of ascorbic acid, an alkaloid and ecliptine (Khare, 2004).

The plant is a good source of thiophene derivatives which are active against nematodes.

The petroleum ether extract of aerial parts contains α-trithienyl aldehyde, ecliptal, besides

stigmasterol and β-sitosterol. The roots are very rich in thiophene acetylenes (Singh, 1988;

Singh and Bhargava, 1992).


E. alba is helpful in reducing the inflammation, pains, early healing of wounds and

also improves the skin texture (Khare, 2004). It is also helpful in improving eyesight,

digestion and is a wonderful liver stimulant (Kirtikar and Basu, 1998). It helps in building

up of the blood cells and maintains the proper density of blood. It is a diuretic agent.

The seeds are aphrodisiac agent. It provides strength to the body. It also helps in

suppressing fever (Khare, 2004).


The herb is used as a tonic and de-obstruent in hepatic and splenic enlargements

and in skin diseases (Mehra and Nanda, 1968). The plant possesses anti-hepatotoxic and

anti-inflammatory activities (Handa et al., 1986).

1.2.16. Literature Pertinent to Argyreia speciosa

Family : Convolvulaceae

English Name : Elephant creeper, Woolly morning glory

Sanskrit Name : Vriddadaru

Hindi Name : Vidhara

Tamil Name : Ambagar, Peymunnai, Sadarbalai

1.2.16.1. Habitat

Argyreia speciosa is found throughout India, upto an altitude of 300 m height

(Kirtikar and Basu, 1995).


A. speciosa is a very large woody climber. The stem is stout, white and tomentose.

The leaves are large, ovate-cordate and glabrous. The flowers are in sub-capitate cymes.

The fruits are globose and apiculate (Kirtikar and Basu, 1995).


Introduction

30 | P a g e


Phytochemical screenings of the plant have shown the presence of ergoline

alkaloids, flavonoids, triterpenes, lipids, tannin and resin (Mann et al., 1999).


The leaves are used by natives as a local stimulant and rubefacient in skin diseases;

the roots are reported to be a tonic, aphrodisiac, bitter, diuretic and used in rheumatism,

gonorrhea, chronic ulcer and diseases of nervous system (Nadkarni, 1976;

Yoganarasimhan, 2000). Pharmacological studies on A. speciosa have been reported it to

possess anti-inflammatory, anti-arthritic (Gokhale et al., 2002), immunomodulatory

(Gokhale et al., 2003), wound healing (Habbu et al., 2007), hepatoprotective activity

(Habbu et al., 2008) and anti-ulcer activity (Abdullah et al., 2010).


The root is bitter, aphrodisiac, diuretic and used in gonorrhoea, rheumatism and

diseases of the nervous system. It is also used as a tonic (Subramoniam et al., 2007;

Krishnaveni and Sent, 2009).

1.2.17. Literature Pertinent to Abhrak Bhasma

English Name : Powered talc, Biotite calx

Sanskrit Name : Abhrak bhasma

Tamil Name : Abhiragam

It is prepared by treating biotite (mica) with the juices of a number of

reconstituents plants that make it a powerful cellular regenerator. It is a nervine tonic and

is also widely used in respiratory tract infection and anemia. It contains iron, magnesium,

potassium, calcium and aluminum in traces (www.himalayahealthcare.com).

1.2.18. Literature Pertinent to Jasad Bhasma

English Name : Zinc calx

Sanskrit Name : Yashad bhasma

Indian Name : Jasad bhasma

Jasad bhasma or Yasad bhasma is one of the members of this class of ayurvedic

pharmaceuticals. It is specially processed zinc. It is administered systemically in diseases


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31 | P a g e

like diabetes, leucorrhoea and hyperhydrosis. The role of the jasad bhasma in arresting

myopia has been examined in one such study (Puri et al., 1983). Zinc as a micronutrient

has been reported as a co-factor of metalloproteinase enzymes like collagenase, which

plays a critical role in wound bed remodeling. More recent studies have shown

unequivocally that topical zinc therapy reduces wound debris and advances

epithelialization in surgical wounds in the rat (Lansdown, 1993; Keefer et al., 1998).

Considering the role of zinc in the wound healing process, the above study was planned to

evaluate the wound healing activity of ayurvedic pharmaceutical product jasad bhasma in

an ointment base for local application (Shah et al., 2009).

1.3. LITERATURE PERTINENT TO HARITAKI

Haritaki is a mother of all herbs. Abhya (which allays the fear of illness), pathya

(beneficial in all diseases) and vyastha (which helps gains longevity), that is how haritaki

has been named in ancient Sanskrit literature (Mahajan and Pai, 2011). Popularly known as

hararh and scientifically as Terminalia chebula, haritaki is one of the oldest herbs known

to mankind. Its moderate-sized deciduous tree is found throughout the greater part of India

and it is the dry pulp of its fruit which is used as medicine (Surya Prakash et al., 2012).

Haritaki fruit is predominantly astringent but at the same time is also bitter, sweet,

pungent and sour in taste and light, dry and hot in effect. It pacifies vata, pitta and kapha,

all the three doshas. Ayurveda has differentiated seven types of haritaki which include

varieties ranging from its different stages of ripeness to the species found according to the

diverse places of its origin. Haritaki has been abundantly praised for its extraordinary

healing properties. Ancient texts have described it to be gentle and caring like a mother

(Gupta et al., 2010).

With a vast array of action on human body, haritaki is primarily digestive,

carminative and laxative in nature. It stimulates liver functions, corrects metabolism, kills

intestinal worms and has tonic effect on all body organs, including the lungs, heart and

brain. Haritaki is also known for its anti-inflammatory, wound healer, anti-obesity,

aphrodisiac and rejuvenating properties (Maruthappan and Sakthi Shree, 2010;

Gupta et al., 2010). In its efficacy and usefulness haritaki has been considered to be equal


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32 | P a g e

to that of amla. The famous Charak Samhita has described it as a drug of choice in the loss

of appetite, indigestion, constipation, upward flow of abdominal gas, sprue and piles.

Besides curing the swelling of liver and spleen, haritaki is also beneficial in cough, asthma,

hiccup, anemia, jaundice, sinusitis and diseases of the urinary tract. Haritaki helps to

dissolve glandular swellings and also has salutary effect in conditions arising due to the

excessive use of alcohol (Jawanjal et al., 2012).

Haritaki is therapeutically prescribed as a preventive and restorative measure.

Though contraindicated in pregnancy and also forbidden for prolonged use, ayurvedic

texts have described different methods to use haritaki in different seasons and diseases

(Mahajan and Pai, 2011). During summer it should be taken with jaggery. In the rainy

season, winter and spring, haritaki is advised to be taken with rock salt, ginger and honey,

respectively. In the diseases arising due to vitiated vata, haritaki should be taken with ghee,

in pitta diseases with sugar and in kapha problems it is indicated to be used with salt

(Jadhav and Laddha, 2004).

As a household remedy, haritaki is best used to clear the bowels. If it is combined

with an equal quantity of amla and baherha a unique combination is achieved which is

known as triphala. Ayurvedic texts have described several uses of triphala, which, besides

being attributed with anti-aging properties, is also given independently or as an adjunct to

cure a number of diseases (Jadhav and Laddha, 2004; Gupta, 2012).

Haritaki habit, habitat, principle constituents, pharmacology and indication are

detailed in section 1.2.15.

1.4. LITERATURE PERTINENT TO AYURSLIM

Ayurslim is an ayurvedic formulation of the pure herbs. Ayurslim is a clinically

proven, safe and effective poly-herbal formulation that actually helps to regulate fat

production and utilization. It also eliminates the craving for sweets, normalizes energy

production and utilization into the body and helps to stay trim and healthy. Ayurslim has a

good effect on weight reduction and lipid profiles (Singh et al., 2008).


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1.4.1. Benefits of Ayurslim

Ayurslim brings about efficient burning of fat. It inhibits fatty acid synthesis,

thereby reducing fat accumulation in the body. It maintains normal cholesterol levels in the

body, factors that are concerned with fat accumulation. It reduces the craving for food and

sweets, thereby reducing the intake of fats and carbohydrates. It brings about

effective utilization of glucose in the body, which has a role to play in fat accumulation in

the body. Ayurslim leads to optimal utilization of nutrients and energy, thereby correcting

energy imbalances in the body that are responsible for fat accumulation. Ayurslim

ingredients include: Garcinia cambogia, Commiphora wightii, Gymnema sylvestre,

Terminalia chebula and Trigonella foenum graecum (www.himalayahealthcare.com).

1.4.2. Contraindications and Ayurslim interactions

No adverse effects have been reported with the use of ayurslim capsules, if taken as

per the prescribed dose. The use of ayurslim capsules is contraindicated in pregnancy,

jaundice and kidney failure. In patients who are already suffering from problems like

diabetes, heart problems and high blood pressure, it is advisable to take ayurslim capsules

under medical supervision. No drug interactions have been reported with the use of

ayurslim capsules (www.himalayahealthcare.com).

1.4.3. Literature Pertinent to Commiphora wightii

Family : Burseraceae

English Name : Indian Bdellium

Sanskrit Name : Guggulu, Koushika, Devadhupa, Palankasha

Hindi Name : Guggul

Tamil Name : Kiluvai, Pachaikiluvai

1.4.3.1. Habitat

Commiphora wightii occurs in the arid rocky tract of Rajputana, Khandesh, Berar,

Mysore, Sind and Baluchistan. C. wightii is a threatened plant species of Indian arid region

(Atal et al., 1975) which is reported from the states of Gujarat and Rajasthan with

restricted distribution. In Gujarat, the species is mainly found in Kachchh and some parts

of Saurashtra regions (Shah, 1978; Soni, 2010).


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C. wightii is a small tree or shrub with spinescent branches. The leaflets are 1to 3 in

number and obovate. The ash coloured bark comes off in rough flakes exposing the under

bark which also peels off in thin papery rolls (Kumar and Shankar, 1982; Soni, 2010).


C. wightii contains an aromatic essential oil besides gum and resin. The gum resin

from the bark contains the octanordammarane terpenes and manusumbionic acid.

The golden yellow oleo-gum-resin is a complex mixture of ketones, several phenolics and

sterols (Bajaj and Dev, 1982), sesquiterpenes (Dolara et al., 2000), guggulsterones

(Swaminathan et al., 1987) and guggutetrols (Kumar and Dev, 1987; Chaudhary, 2012).


It is used in the allopathic, ayurvedic and unani systems of medicines due to its

anti-inflammatory, anti-rheumatic, hypocholesteremic, hypolipidemic and anti-fertility

activities (Tajuddin et al., 1997). The oleo-gum resin of C. wightii is an efficacious

treatment for arthritis, obesity, bone fractures, inflammation, cardiovascular disease and

lipid disorders (Dev, 1997; Singh et al., 2003; Soni, 2010).


C. wightii acts as a bitter and carminative, stimulating the appetite and improving

digestion. It causes an increase in leucocytes in the blood and stimulates phagocytosis. The

resin is used in the form of a lotion for indolent ulcers and as a gargle in chronic tonsillitis,

pharyngitis and ulcerated throat (Arora et al., 1971; Singh et al., 2003).

1.4.4. Literature Pertinent to Garcinia cambogia

Family : Guttifererae

English Name : Garcinia

Sanskrit Name : Vrikshamia, Kankusta

Tamil Name : Kodukkaippuli

1.4.4.1. Habitat

Garcinia cambogia is commonly found in the evergreen and shoal forests of

Western Ghats in India up to 6,000 ft height (Hammer, 2001).


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G. cambogia is a small or medium sized tree with a rounded crown and horizontal

or drooping branches. The leaves are dark green shining elliptic to obovate. Fruits are

ovoid and yellow when ripe, with 6 to 8 grooves. Seeds are succulent aril (Hammer, 2001).


G. cambogia contains essentially hydroxy-citric acid (Jayaprakasha and Sakariah,

1998; Revathi et al., 2010), camboginol and cambogin (Sriyani and Guntalika, 1998).


Garcinia extract inhibits the cytoplasmic lipid accumulation as well as adipogenic

differentiation of pre-adipocytes (Kim et al., 2004). Garcinia cambogia extract is a herbal

preparation that has been suggested as a useful in the treatment of gastrointestinal

disorders (Mahendran et al., 2002), anti-obesity activity (Shara et al., 2004;

Preuss et al., 2005; Kim et al., 2008), anti-cancer activity (Liao et al., 2005),

anti-inflammatory activity (Dos Reis et al., 2008), lipid lowering property (Asha Koshy

and Vijayalakshmi, 2001) and anti-helminthic activity (Mathew et al., 2011).


In ayurveda, it is used as sour flavour and activates digestion. This herb has been

used historically in India to supsport the treatment of various health conditions

(Clouatre and Rosenbaum, 1994).

1.4.5. Literature Pertinent to Terminalia chebula

As detailed in section 1.2.15.

1.4.6. Literature Pertinent to Trigonella foenum-graecum

Family : Fabaceae

English Name : Fenugreek

Sanskrit Name : Medhika, Chandrika

Tamil Name : Venthayam

1.4.6.1. Habitat

Trigonella foenum graecum is an aromatic, 30 to 60 cm tall, annual herb and

cultivated throughout the country (Kirtikar and Basu, 1935).


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1.4.6.2. Morphological Description

It is a nearly smooth erect annual. Leaflets are 2 to 2.5 cm long, oblong and

toothed. Flowers are 1 to 2 auxiliary and sessile. Pod is 5 to 7.5 cm long, 10 to 29 seeded

and without transverse reticulations (Kirtikar and Basu, 1935).


Fenugreek is a rich source of steroidal sapogenins (Hardman, 1969). Fenugreek

seed contains carbohydrates, proteins lipids, pryridine type alkaloids mostly trigonelline,

choline, gentianine, carpaine, flavonoids, calcium, iron, saponins, glycosides, sitosterol,

vitamins A, B1, C, nicotinic acid and volatile oils (Shang et al., 1998).


Fenugreek seeds are considered carminative, tonic and aphrodisiac and used in

dyspepsia with loss of appetite and in rheumatism (Kirtikar and Basu, 1935). The seeds are

hot and dry; suppurative, diuretic, useful in dropsy, chronic cough, enlargement of the liver

and the spleen. The leaves are useful in external and internal swellings and burns; prevent

the hair falling off (Kirtikar and Basu, 1935). It is also considered to be hypoglycemic

(Jain et al., 1987) and anti-fertility agent (Kamal et al., 1993).


Fenugreek seeds are used as a traditional remedy for the treatment of diabetes

(Miraldi et al., 2001; Basch et al., 2003), dysentery, diarrhoea, inflammatory colic and as

an anti-bacterial (Omolosa and Vagi, 2001).

1.4.7. Literature Pertinent to Gymnema sylvestre

Family : Asclepiadaceae

English Name : Gymnema

Sanskrit Name : Meshashringi, Vishani, Madhunashini

Hindi Name : Gurmar, Merasingi

Tamil Name : Sirukurinjan

1.4.7.1. Habitat

Gymnema sylvestre is found in the Deccan Peninsula, extending to parts of

Northern and Western India (Kirtikar and Basu, 1975; Grover et al., 2002).


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G. sylvestre is a large, more or less pubescent and woody climber. The leaves are

opposite and ovate; the flowers are small, yellow and in umbellate cymes (Kirtikar and

Basu, 1975; Zhen et al., 2001).


G. sylvestre constituents include resins, gymnemic acids, saponins, stigmasterol,

quercitol and the amino acid derivatives betaine, choline and trimethylamine

(Shah et al., 2010), albumin, carbohydrates, tartaric acid, formic acid, butyric acid, inositol

alkaloids, parabin, calcium oxalate, lignin and cellulose (Sinsheimer and Rao, 1970).


It is bitter, astringent, acrid, thermogenic, anti-inflammatory, anodyne, digestive,

liver tonic, emetic, diuretic, stomachic, stimulant, anti-helminthic, alexipharmic, laxative,

cardiotonic, expectorant, anti-pyretic and uterine tonic. It is useful in inflammations,

hepatosplenomegaly, dyspepsia, constipation, jaundice, haemorrhoids, strangury renal and

vesicle calculi, helminthiasis, cardiopathy, cough asthma, bronchitis, intermittent fever,

amenorrhoea, conjunctivitis and leucoderma (Chopra et al., 1992).


Gymnemic acid has anti-diabetic property. It has an inhibitory effect on plasma

glucose and serum insulin in man. The plant is stomachic, stimulant, laxative, diuretic,

anti-obesity, anti-microbial activity and anti-hyperglycemic activity (Baskaran et al., 1990;

Shah et al., 2010).

1.5. LITERATURE PERTINENT TO HORMONAL PARAMETERS

1.5.1. Literature Pertinent to Thyroid Hormones

Some environmental chemicals can disrupt the thyroid hormone, which is essential

for the development of the brain and other organs. It is also important for the general

functioning of the body. Thyroid hormones control the body’s metabolism (lethargy, body

temperature and weight). The thyroid gland requires iodine to produce the two main

thyroid hormones, T4 and T3. Thyroid hormones travel in the bloodstream and then

enter cells. They work by binding to thyroid receptors (TRs) in the cell nucleus,

where they perform some specific function and stimulates cell metabolism (Zoeller, 2005;


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38 | P a g e

Rajender et al., 2011). The pituitary gland releases thyroid stimulating hormone, or TSH,

to prompt the thyroid gland to produce more thyroid hormone. If the pituitary perceives

there is not enough thyroid hormone in the body, TSH levels rise. Conversely,

when the thyroid gland releases T4, it suppresses the pituitary’s further release of TSH

(Patandin et al., 1999; Zoeller, 2005).

1.5.1.1. Mechanism of Action

Thyroid hormone (T3 and T4), produced by the thyroid gland, plays an important

role in development, differentiation and metabolism (Lazar, 1993). The lack of T3 and

T4 in early human development results in growth disturbances and severe mental

retardation, a disease called cretinism (Oppenheimer, 1983). T3 action is mediated by

nuclear T3 receptors (TRs) that can bind T3 with high affinity (Lazar, 1993). TRs belong to

the nuclear receptor super family that also includes the receptors for retinoids, vitamin D,

fatty acids and prostaglandins (Evans, 1988; Glass, 1994; Ribeiro et al., 1995). Thyroid

hormone works through the same general mechanism as steroid hormones. It first binds to

cytoplasmic and/or nuclear receptors which once activated will migrate to the nucleus to

regulate the transcription of specific genes (Zhang and Lazar, 2000; Harvey and

Williams, 2002).

1.5.1.2. Thyroid Hormone and Obesity

In obese subjects, decreasing weight loss with continued caloric deprivation

(Moore et al., 1980) has been attributed to reduce serum T3 concentrations inducing a

lower metabolic rate (Moreira-Andres et al., 1981). Consequently, T3 or T4, in varying

doses and duration, has been administered to euthyroid obese subjects during caloric

deprivation to enhance weight loss (Adler and Wartofsky, 2007). Likewise, euthyroid

patients with non-thyroidal disorders have been treated with T3 and/or T4 in an attempt to

improve morbidity and mortality (Burman et al., 1979; Adler and Wartofsky, 2007).

1.5.1.3. Metabolism

Thyroid hormones stimulate diverse metabolic activities in most tissues, leading to

an increase in basal metabolic rate. One consequence of this activity is to increase body

heat production, which seems to result, at least in part, from increased oxygen

consumption and rates of ATP hydrolysis (Bianco et al., 2002).


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1.5.1.3.1. Effects on Growth and Metabolism

Tri-iodothyronine, the active form of thyroid hormone, acts on receptors in nearly

every type of cell in the body to activate coding for a number of genes. The result is an

increased production of proteins-proteins that act as enzymes, proteins required for tissue

growth and those needed for transport of other substances. This leads to increased

metabolic activity in the body. Large quantities of the hormone can raise basal metabolism

rate (Fisher et al., 1982).

1.5.1.3.2. Effects on weight Maintenance of weight gain or loss is associated with compensatory changes

in energy expenditure that oppose the maintenance of a body weight that is different from

the usual weight (Leibel et al., 1995; Rosenbaum et al., 1996). Catecholamine release

in response to insulin-induced hypoglycemia is diminished in reduced obese

patients (Jung et al., 1982; Landsberg and Young, 1984; Leibel et al., 1991); serum

tri-iodothyronine decreases in subjects during the process of weight loss and increases in

subjects during dynamic weight gain (Danforth and Burger, 1984). The decreased insulin

sensitivity after weight gains the beneficial effects of even modest amounts of weight

reduction on carbohydrate metabolism (Kolterman et al., 1980; Jimenez et al., 1987).

1.5.1.3.3. Effect on Protein Metabolism

The amino acids required for continued hepatic gluconeogenesis are also derived

from proteolysis, particularly muscle proteins. Thyroid hormones also increase protein

synthesis but the degradation of protein usually outweighs the synthesis so there is still net

loss of muscle proteins. This is particularly observed in cases of excessive

tri-iodothyronine production wherein muscle wasting and weakness occur as well as

increased nitrogen loss through the urine as urea (Dumitrescu and Refetoff, 2011).

1.5.1.3.4. Effect on Carbohydrate Metabolism

All aspects of carbohydrate metabolism are increased by thyroid influence,

including glucose production by the liver particularly through gluconeogenesis.

Gluconeogenesis is the production of glucose by the liver from stored fats or proteins.

This effect does not increase plasma glucose concentrations because the pancreas is also

stimulated by the hormone to secrete increased amounts of insulin to keep up with

increased glucose production (Potenza et al., 2009).


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1.5.1.3.5. Effect on Fat and Lipid Metabolism

Increased thyroid hormone levels stimulate fat mobilization, leading to increased

concentrations of fatty acids in plasma. They also enhance oxidation of fatty acids in many

tissues. Plasma concentrations of cholesterol and triglycerides are inversely correlated with

thyroid hormone levels. The diagnostic induction of hypothyroidism is increased blood

cholesterol concentration (Thompson et al., 1981; Ness et al., 1990; Rizos et al., 2011).

1.5.1.3.6. Effect on Insulin Secretion

It is well established that thyroid hormones affect insulin action (Dimitriadis and

Raptis, 2001). Thyroid hormone excess has been consistently found to induce insulin

resistance (Dimitriadis et al., 1985; Dimitriadis and Raptis, 2001). Glucose regulation by

insulin depends on the suppression of endogenous glucose production and the

stimulation of peripheral glucose disposal. Hepatic glucose production is decreased

(Okajima and Ui, 1979; McCulloch et al., 1983) or unchanged (Muller et al., 1988) in

hypothyroidism, but there is little information on the effects of insulin in peripheral tissues.

1.5.2. Literature Pertinent to Glucocorticoids

Glucocorticoids are also known as cortisol and produced in the adrenal cortex.

It controls the synthesis of fats, proteins and carbohydrates, which, together with cortisol,

suppress inflammatory reactions in the body and also affects the immune system.

Glucocorticoids are used to stop the inflammation process. The inflammatory process has

evolved in the body for a useful purpose; namely as a defensive reaction to the damage or

injury to tissue. By a series of reactions, inflammation is designed to isolate whatever is

causing the irritation, help eradicate the presumed invader and help repair the surrounding

damaged tissue (Necela and Cidlowski, 2004; Rhen and Cidlowski, 2005).

1.5.2.1. Mechanism of Action

Glucocorticoids penetrate their target cell membranes and binds to intracellular

receptors. The intracellular mediator, synthesized in response to genome stimulation by

cortisol is the protein lipocortin-1 (LC-1). Glucocorticoids on binding to glucocorticoid

receptors promote preferential transcription of certain DNA segments and this leads

via the appropriate mRNAs to the synthesis of enzymes that alter cell function

(Dallman et al., 1993).


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1.5.2.2. Cortisol in Liver

The liver is a major target for glucocorticoids; the effects on the liver leading to

insulin resistance likely affect the entire body. The measurements of glucocorticoid

metabolism for the body as a whole usually reflect the activity in the liver

(Tomlinson and Stewart, 2005). The high levels of glucocorticoids in visceral obesity

increase the activity of lipoprotein lipase (LPL), so lipoprotein triglycerides are converted

to free fatty acids. Subcutaneous adipocytes are more sensitive to insulin, but visceral

adipocytes are more sensitive to glucocorticoid stimulation. Therefore, lipolysis of fat,

inhibited by insulin, is more pronounced in visceral fat than in subcutaneous fat

(Goldstein, 2002). This increases the release of free fatty acids (FFA) in visceral adipose

tissue (VAT); because VAT drains into the portal vein, this may increase the free

fatty acids, glucocorticoids and adipokines that reach the liver through the blood and

contribute to hepatic insulin resistance (Samaras and Campbell, 1997; Rosmond, 2003).

1.5.2.3. Cortisol in Muscle and Pancreas

In myoblasts, increased expression of 11β-HSD1 and the glucocorticoid

receptor (GR) in muscle are correlated with increased insulin resistance and BMI

(Stulnig and Waldhausl, 2004). The high glucocorticoid levels inhibit glycogen synthase

and promote the release of free fatty acids. Additionally, the glucocorticoids in muscles

may promote the expression of fatty acid transporter genes (Qi and Rodrigues, 2007).

Therefore, triglycerides may collect in the muscles and muscles may be affected with

hyperinsulinemia (Rosmond, 2003; Abate and Chandalia, 2003). Glucocorticoids likely act

on downstream insulin signaling in the muscle and therefore decrease the uptake of

glucose, contributing to insulin resistance and type II diabetes (Qi and Rodrigues, 2007).

In the pancreas, cortisol can impair insulin-dependent uptake of glucose.

Also, increased cortisol produced by increased 11β-HSD1 action in islet cells may hinder

the secretion of insulin from β-cells of the pancreas (Stulnig and Waldhausl, 2004).

1.5.2.4. Glucocorticoids and Obesity

Obesity is a prevalent condition and is associated with premature mortality

from vascular disease. For any given body mass index (BMI), mortality is higher if

fat is distributed centrally (visceral adiposity) compared with a more generalized

pattern of distribution (Bjorntorp, 1997). Glucocorticoids appear to be one such factor.


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Patients with Cushing’s syndrome develop central obesity, which improves with resolution

of the hypercortisolism (Mayo-Smith et al., 1989). On this basis, several studies have

evaluated the hypothalamic-pituitary-adrenal axis in patients with obesity, but these have

invariably focused on circulating and urinary concentrations and secretion rate. Overall,

the circulating cortisol concentrations are normal in patients with obesity (independent of

adipose tissue distribution); secretion rates are higher, particularly in patients with visceral

adiposity (Hautenen and Adlercreutz, 1993; Ljung et al., 1996).

1.5.2.5. Effects on Metabolism

In the fasted, state cortisol stimulates several processes that collectively serve to

increase and maintain normal concentrations of glucose in blood. These effects include

stimulation of gluconeogenesis, particularly in the liver, mobilization of amino acids form

extra hepatic tissues, inhibition of glucose uptake in muscle and adipose tissue and

stimulation of fat breakdown in adipose tissue i.e., fatty acids released by lipolysis

(Friedman et al., 1993; Argaud et al., 1996).

1.5.3. Literature Pertinent to Insulin

Insulin is synthesized as a pre-prohormone in the β-cells of the islets of langerhans.

Insulin secretion from β-cells is principally regulated by plasma glucose levels. Increased

uptake of glucose by pancreatic β-cells leads to a concomitant increase in metabolism.

The increase in metabolism leads to an elevation in the ATP/ADP ratio. This in turn leads

to the inhibition of an ATP-sensitive potassium channel (KATP channel). The net result is a

depolarization of the cell leading to Ca2+

influx and insulin secretion (Huopio et al., 2002;

Geng et al., 2003).

The major function of insulin is to counter the concerted action of a number of

hyperglycemia generating hormones and to maintain low blood glucose levels. Because

there are numerous hyperglycemic hormones, untreated disorders associated with insulin

generally lead to severe hyperglycemia (Fowelin et al., 1993). In addition to its role in

regulating glucose metabolism, diminishes lipolysis and increases amino acid transport

into cells. Insulin also modulates transcription, altering the cell content of numerous

mRNAs. It stimulates growth, DNA synthesis and cell replication, effects that it holds in

common with the insulin like growth factors (IGFs) and relaxin (Kersten, 2001).


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1.5.3.1. Role of Insulin in the Fat Cells

Fat synthesis and storage are facilitated by a rise in the plasma level of insulin.

A decrease in the plasma level of insulin facilitates the breakdown of stored fat, making it

available for oxidation by the muscle cells and numerous other body cells. A decrease in

the insulin level enhances the breakdown; the release and the burning of stored fat

(Cohen et al., 1996).

Hormone sensitive lipase (HSL) is the key enzyme in the metabolic pathway in

the fat cell that is influenced by insulin to produce its anabolic and catabolic effects on

fat. When the plasma level of insulin is elevated, HSL is inhibited and the anabolic effect

of insulin occurs. Fatty acids move into the fat cells and triglycerides are formed and

stored. When insulin level is lowered, its inhibitory effect on HSL is removed and the

catabolic effect of insulin results. Triglycerides are broken down and fatty acids leave the

fat cells and become available for oxidation by various cells of the body

(Makino et al., 1992; Habener et al., 1999; Lim et al., 2011).

1.5.3.2. Role of Insulin in the Liver and Muscle Cells

An increased plasma level of insulin facilitates the transport of glucose into various

cells, including the muscle cells, the primary site of energy utilization. In the liver,

elevation in insulin level stimulates the synthesis of glycogen. A reduction in the plasma

insulin level stimulates the synthesis of glucose (primarily in the liver) from metabolites

such as lactate, pyruvate, oxaloacetic acid, alanine, leucine and numerous other amino

acids (gluconeogenesis) (Malaisse et al., 1998; Swithers and Davidson, 2010).

1.5.3.3. Insulin and Obesity

In case of obesity, the fat accumulation in the adipose tissue increases.

Excess abdominal adipose tissue has been shown to release increased amount of free fatty

acids which directly affect insulin signaling, diminish glucose uptake in muscle, drive

exaggerated glucose synthesis and induce gluconeogenesis in liver. A number of

mechanisms were proposed to explain the development of insulin resistance caused by

elevated free fatty acids. Free fatty acids released by the visceral adipose tissue enter the

portal vein and reach the liver. In the liver they interact with the hepatocytes and immune

cells. This finally leads to insulin resistance and decrease in glucose uptake by hepatocytes

as well as increase in the production of glucose (Girard and Lafontan, 2008).


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1.5.3.4. Insulin and Carbohydrate Metabolism

Glucose is liberated from dietary carbohydrate such as starch or sucrose by

hydrolysis within the small intestine and is then absorbed into the blood.

Elevated concentrations of glucose in blood stimulate release of insulin and insulin acts on

cells throughout the body to stimulate uptake, utilization and storage of glucose.

The effects of insulin on glucose metabolism vary depending on the target tissue

(Davidson and Berliner, 1972; Bhatt, 2010).

1.5.3.5. Insulin and Free Fatty Acids

Fatty acids are synthesized by the extra mitochondrial system which is responsible

for the complete synthesis of palmitate from acetyl-CoA in the cytosol. This system is

present in many tissues, including liver, kidney, brain, lung, mammary gland and adipose

tissue. Although the main role of fatty acids is to reserve energy, they play a significant

role in insulin utilization by liver and muscle and glucose stimulated insulin secretion

(GSIS) from pancreas through GPR40 (Boden and Shulaman, 2002). Aberration in the

process of fatty acid oxidation leads to diseases associated with hypoglycemia.

Elevated levels of free fatty acids which are not bound to plasma albumin play an

important role in development of insulin resistance and impairment of β cell function,

which are the main causes of hyperglycemia. It is important to distinguish between insulin

resistance in adipose tissue and subsequent elevation of plasma fatty acids and

mechanisms of free fatty acid induced insulin resistance (Michael and Peter, 2005;

Tuei et al., 2011).

1.5.3.6. Insulin and Lipid Metabolism

Insulin inhibits breakdown of fat in adipose tissue by inhibiting the intracellular

lipase that hydrolyzes triglycerides to release fatty acids. Insulin facilitates entry of glucose

into adipocytes and within those cells; glucose can be used to synthesize glycerol.

This glycerol, along with the fatty acids delivered from the liver, is used to synthesize

triglyceride within the adipocyte. By these mechanisms, insulin is involved in further

accumulation of triglyceride in fat cells (Moller, 2001; Chang et al., 2011).


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1.6. LITERATURE PERTINENT TO HAEMATOLOGICAL PARAMETERS

Blood is a specialized bodily fluid that delivers necessary substances to the body's

cells (in animals) such as nutrients and oxygen and transports waste products away from

those same cells. In vertebrates, it is composed of blood cells suspended in a liquid called

blood plasma. Plasma, which constitutes 55% of blood fluid, is mostly water (92% by

volume) and contains dissipated proteins, glucose, mineral ions, hormones, carbon dioxide

(plasma being the main medium for excretory product transportation), platelets and blood

cells themselves. The blood cells are mainly red blood cells (also called RBCs or

erythrocytes), white blood cells and platelets. Full blood count is a frequently used

laboratory test performed to support the diagnosis of several diseases; anemia,

certain cancers, infections, acute hemorrhagic states, allergies and immunodeficiency

disorders or used in periodic health examination and preoperative evaluation

(George and Parker, 2003).

1.6.1. Literature Pertinent to Erythrocytes or Red Blood Corpuscles

Erythrocytes contain haemoglobin, which is the colour source of the blood.

The role of erythrocytes is the transport of oxygen and carbon dioxide and is very flexible.

The normal life span of RBC is about 120 days, during that time 85% cells would be

engulfed by the macrocyte-macrophage (Fitzgerald, 1999). The remaining 15% of RBC’s

undergo haemolysis in circulation (Herdy, 1996). The presence of chronic infection can

shorten the life span of the RBC. In addition, conditions causing chronic inflammation,

such as rheumatoid arthritis, cancer and liver diseases can truncate the RBC lifespan

(Linker, 1996).

1.6.1.1. Erythrocytes and Obesity

Obesity is associated with increased tendency of RBC to adhere to each other and

form aggregates (Samocha-Bonet et al., 2003). RBC aggregation depends on opposing

factors: the repulsive force between the negatively charged cells and the disaggregating

shear force generated by blood flow, on the one hand and the cell to cell adhesion induced

by plasma proteins, on the other (Chien, 1982; Nash et al., 1987). Thus, RBC aggregation

is dependent on both cellular (intrinsic) and plasmatic (extrinsic) factors

(Samocha-Bonet et al., 2003).


http://en.wikipedia.org/wiki/Cell_(biology)

http://en.wikipedia.org/wiki/Animal

http://en.wikipedia.org/wiki/Waste

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Inflammation is characterized by an enhanced synthesis of adhesive proteins

including fibrinogen (De Maat et al., 1999), gamma globulins (Imaizumi and Shiga, 1983),

CRP (Gabay, 1999) and other macromolecules that might influence the aggregability

of red blood cells (RBCs) (Dalakas, 1994; Weng et al., 1998). Enhanced RBC aggregation

has been consistently reported in various cardiovascular risk factors including

hyperlipoproteinemia (Vaya et al., 1993; Weng et al., 1998), diabetes mellitus

(Caimi et al., 1993) and hypertension (Ravazian et al., 1992). Obesity has also been

reported to influence erythrocyte rheological properties, probably through

hyperinsulinemia and glucose intolerance (Valensi et al., 1996).

In addition to conventional cardiovascular risk factors associated to obesity, which

could partly explain the increase in thrombotic risk, it has been suggested that

haemo-rheological disturbances, such as an increase in erythrocyte aggregability (EA)

(Lowe et al., 2000; Brun et al., 2004; Sola et al., 2007) or a decrease in erythrocyte

deformability (ED) (Solerte et al., 1997; Perez-Martin et al., 2001) may favor the

development of thrombotic events.

1.6.2. Literature Pertinent to Leucocytes or White Blood Corpuscles

White blood cells are the cellular part of the immune system and are very important

in surveying the body for infection. They find, trap, neutralize and kill invading pathogens.

There are different types of WBCs, which have specific functions in protecting against

developing infections (Gleeson, 2007). WBCs are nucleated non-pigmented or colourless

cells described as wandering cells and are not confined to the blood channel. An important

function of WBC is phagocytosis (Prakash and Arora, 1998). They are present in blood in

considerable fewer number than the RBC. These are usually one per every 500 RBCs

(Carman, 1993).

1.6.2.1. Leucocytes and Obesity

The adipocyte is an important source of cytokines, namely interleukin (IL)-6 and

tumour necrosis factor (TNF)-α and their levels are significantly higher in the plasma of

obese patients (Berg and Scherer, 2005; Rondinone, 2006). The rise in these cytokines,

especially in IL-6, triggers an increased synthesis of C-reactive protein (CRP), one of the

most sensitive makers of inflammation. However, obesity is recognized as a possible cause

for reactive leucocytosis (Herishanu et al., 2006; Veronelli et al., 2011).


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47 | P a g e

1.6.2.2. Effect on Immune System

WBCs are necessary for protection against invading organisms and protection of

the immune system and are markers of inflammation. An elevated white blood cell count is

a predictor of cardiovascular mortality independent of the effects of smoking and other

traditional risk factors (Gillum et al., 1993; Weijenberg et al., 1996; Brown et al., 2001).

There is also a significant positive association between the WBC count and the severity of

carotid atherosclerosis (Elkind et al., 2001). Inflammation contributes to vascular injury,

atherogenesis and thrombosis (Mehta et al., 1998; Ross, 1999). WBC, which is activated

by cytokines, especially IL-6 and IL-8 (Van Oostrom et al., 2003), may serve as an

important marker of these processes (Ernst et al., 1987; De Servi et al., 1991).

WBCs contribute to blood viscosity, release products that induce plaque rupture and

thrombus formation (Ernst et al., 1987) and have a role in endothelial dysfunction

(Murohara et al., 1994).

1.6.2.3. Effect on Glucose Metabolism

Impaired glucose tolerance (IGT) is often associated with the metabolic syndrome

and is an established risk factor for cardiovascular disease (Tominaga et al., 1999).

White blood cell (WBC) count is elevated in obesity (Kullo et al., 2001) and is a risk

factor for atherosclerosis (Elkind et al., 2001). An elevated WBC count is present in

impaired glucose tolerance (IGT) (Ohshita et al., 2004; Veronelli et al., 2011) and WBC

count is associated with macro and micro-angiopathic complications in type II diabetes

(Tong et al., 2004).

1.6.3. Literature Pertinent to Haemoglobin

Haemoglobin is a red protein pigment and the main component of RBC that is

responsible for transporting oxygen from the lungs to tissues for energy. It also carries

carbon dioxide from the tissues to the lungs for excretion. The haemoglobin molecule

consists of two parts: a porphyrin group or haeme, and the protein or globin portion.

Globin is made up of four polypeptide chains attached to the porphyrin ring. In the normal

subject these chains can be of four types: alpha, beta, delta and gamma. In normal and

abnormal haemoglobins (with the exception of haemoglobin H and Bart’s), two sets of

identical polypeptide chains make up the globin (Gleeson, 2007).


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Iron deficiency remains the most common nutritional deficiency in the world.

Iron deficiency has been linked to behavioral and learning problems among children

(Lozoff et al., 2000; Pollitt, 2001) and adolescents (Halterman et al., 2001), increased risks

for preterm infants and small infants among pregnant women (Rasmussen, 2001) and

problems with work and exercise capacity among adults (Baynes and Bothwell, 1990).

1.6.4. Literature Pertinent to Mean Corpuscular Haemoglobin (MCH)

The MCH denote the average haemoglobin weight per red blood cell and is

calculated by multiplying the haemoglobin in gram/100 ml of blood by 10 and dividing the

product by the erythrocyte count in millions per cubic millimeter. It is a calculation of the

amount of oxygen carrying haemoglobin inside the RBCs, since macrocytic RBCs are

larger than either normal or microcytic RBCs, they would also tend to have higher MCH

values (Mbaka et al., 2010).

1. 7. LITERATURE PERTINENT TO BIOCHEMICAL PARAMETERS

1.7.1. Literature Pertinent to Total Proteins

Proteins contain carbon, hydrogen, oxygen, nitrogen and sometimes other atoms.

They form the cellular structural elements, are biochemical catalysts and are important

regulators of gene expression. Nitrogen is essential to the formation of twenty different

amino acids; the building blocks of all body cells. Amino acids are characterized by the

presence of a terminal carboxyl group and an amino group in the alpha position and they

are connected by peptide bonds (Murray et al., 1996; Bland et al., 1999).

Digestion breaks protein down to amino acids. If amino acids are in excess of the

body's biological requirements, they are metabolized to glycogen or fat and subsequently

used for energy metabolism. If amino acids are to be used for energy their carbon

skeletons are converted to acetyl CoA, which enters the Krebs cycle for oxidation,

producing ATP. The final products of protein catabolism include carbon dioxide, water,

ATP, urea and ammonia (Ekhard and Filer, 1996; Metges and Barth, 2000).

Body proteins are broken down when dietary supply of energy is inadequate during

illness or prolonged starvation. The proteins in the liver are utilized in preference to those

of other tissues such as the brain. The gluconeogenesis pathway is present only in liver

cells and in certain kidney cells (Fenuku, 1982; Brinkworth et al., 2004). Total protein


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may be elevated due to chronic infection, adrenal cortical hypofunction, liver dysfunction,

collagen vascular disease (rheumatoid arthritis, systemic lupus and scleroderma),

hypersensitivity states, sarcoidosis, respiratory distress, haemolysis, cryoglobulinemia,

alcoholism and leukemia. Total protein may be decreased due to: malnutrition and

malabsorption (insufficient intake and/or digestion of proteins), liver disease (insufficient

production of proteins), diarrhoea (loss of protein through the GI tract), severe burns

(loss of protein through the skin), hormone imbalances that favor breakdown of tissue,

loss through the urine in severe kidney disease (proteinuria), low albumin, low globulins

and pregnancy (dilution of protein due to extra fluid held in the vascular system)

(Fenuku, 1982; Skov et al., 1999).

1.7.1.1. Protein Metabolism and Obesity

Body fat distribution has been demonstrated to be an important variable in

predicting the metabolic abnormalities accompanying human obesity. Upper body obesity

is more likely to result in hyperlipidemia, diabetes and hypertension (Evans et al., 1984).

Resistance to insulin’s glucoregulatory effects are more pronounced in upper body

obesity than in lower body obesity (Evans et al., 1984a) and specific differences

in effective adipose tissue lipolysis (Jensen et al., 1989). Other differences include greater

peripheral hyperinsulinemia in upper body obesity, most likely a result of reduced hepatic

insulin extraction (Peiris et al., 1986) and different proportions of fast-twitch (type II)

and slow twitch (type I) muscle fiber types (Lillioja et al., 1987). It is not known

whether differences in body fat distribution in human obesity are also associated with

differences in regulation of protein metabolism; however, insulin (Fukagawa et al., 1985),

FFA availability (Tessari et al., 1986) and muscle fiber type (Flaim et al., 1980) are known

to influence protein metabolism.

1.7.1.2. Thermogenic Effects of Proteins

The thermic effect of nutrients is related to the stimulation of energy requiring

processes during the post prandial period. It is based on the amount of ATP required for

the initial steps of metabolism and storage. Thus, a high protein diet induces a greater

thermic response in healthy subjects compared to a high fat diet (Westerterp et al., 1999).

This even implied a higher fat oxidation, thus a negative fat balance and a positive protein

balance (Lejeune et al., 2006).


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1.7.1.3. Gluconeogenesis

The main gluconeogenic organ is the liver. The activity of hepatic

phosphoenolpyruvate carboxykinase (PEPCK), an enzyme involved in gluconeogenesis, is

increased in rats fed a high protein diet. This effect is observed with diet containing protein

or not carbohydrates suggesting that the level of protein in the diet is able to stimulate

hepatic gluconeogenesis (Peret and Chanez, 1975; Bois-Joyeux and Chanez, 1986).

Liver gluconeogenesis is stimulated by a high protein diet but that in the fed state the

newly synthesized glucose-6-phosphate (G-6-P) is directed toward glycogen synthesis

whereas in the fasted state it is converted to glucose and released from hepatocyte.

The control of PEPCK and G-6-Pase activity in the liver by nutrients has a profound

impact on hepatic metabolism and glucose homeostasis and the satiating effect of high

protein feeding could be related to the improvement of glucose homeostasis through the

modulation of hepatic gluconeogenesis and subsequent glucose metabolism

(Seoane and Trinh, 1997; Trinh and O’Doherty, 1998).

1.7.1.4. Insulin Sensitivity

The insulin stimulating effect of proteins may be mediated through specific amino

acids released during digestion. Several amino acids are potent stimulators of insulin

release and certain amino acids (e.g., leucine, arginine, phenylalanine and tyrosine) are

more insulinogenic than are others (Van Loon et al., 2000; Calbet and MacLean, 2002).

Aerobic and/or resistance exercise increase insulin sensitivity as well as the ability of

protein/amino acid intake to stimulate muscle anabolism (Biolo et al., 1997; 1999).

Exercise accelerates muscle protein turnover; however, stimulation of protein synthesis

exceeds activation of proteolysis (Biolo et al., 1995).

1.7.2. Literature Pertinent to Glucose

Carbohydrates can serve as energy source for animals. The breakdown of organic

constituents mainly carbohydrates has a vital role in energy yielding process. The major

function of carbohydrates in metabolism is to provide energy for cellular activities.

A variety of enzyme systems are associated either directly or indirectly with the

metabolism of carbohydrates in many pathological conditions. The metabolism of

carbohydrates gained an importance in the overall physiology of an animal and this forms

an evidence to understand the biochemical state of the cell (Jeffrey et al., 1999).


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1.7.2.1. Carbohydrate Metabolism

Carbohydrate metabolism plays an important role in both types of diabetes

mellitus. The entry of glucose into most tissues including heart, muscle and adipose tissue

is dependent upon the presence of the hormone insulin. Insulin controls the uptake and

metabolism of glucose in these cells and plays a major role in regulating the blood glucose

concentration. The reactions of carbohydrate metabolism cannot take place without the

presence of the vitamin B, which function as coenzymes (Murray et al., 1996;

Jeffrey et al., 1999).

Carbohydrate metabolism begins with glycolysis, which releases energy from

glucose or glycogen to form two molecules of pyruvate, which enter the Krebs cycle, an

oxygen requiring process, through which they are completely oxidized. Before the Krebs

cycle can begin, pyruvate loses a carbon dioxide group to form acetyl coenzyme-A

(acetyl CoA). This reaction is irreversible and has important metabolic consequences.

The conversion of pyruvate to acetyl CoA requires the vitamin B (Linder, 1991;

Shils et al., 2006).

Glycogenesis is the conversion of excess glucose to glycogen. Glycogenolysis is

the conversion of glycogen to glucose (which could occur several hours after a meal or

overnight) in the liver or, in the absence of glucose-6-phosphate in the muscle, to lactate.

Gluconeogenesis is the formation of glucose from non-carbohydrate sources, such as

certain amino acids and the glycerol fraction of fats when carbohydrate intake is limited.

Liver is the main site for gluconeogenesis, except during starvation, when the kidney

becomes important in the process. Disorders of carbohydrate metabolism include diabetes

mellitus, lactose intolerance and galactosemia (Salway, 1999; Wardlaw et al., 2002).

1.7.2.2. Carbohydrate and Obesity

A systematic review of low carbohydrate diets found that the weight loss is

associated with the duration of the diet and restriction of energy intake, but not with

restriction of carbohydrates (Astrup et al., 2004). Although many environmental factors

promote a positive energy balance, it is clear that the consumption of a low carbohydrate,

high fat diet increases the likelihood of weight gain (Saris, 2003). Certainly, many studies

have demonstrated the beneficial effects of high carbohydrate, low fat diets to reduce

adiposity and other aspects of the metabolic syndrome (Schroder et al., 2004).


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The high fat diets produce a more metabolically efficient state may be that glucose

produces more post prandial thermogenesis than fats (Westerterp, 2004). Specifically,

increasing plasma glucose either by infusion or ingestion (Rothwell et al., 1983) increases

metabolic rate. However, per calorie, carbohydrates produce about three fold more

thermogenesis than fat (Almind and Kahn, 2004; Westerterp, 2004). At least part of

glucose induced thermogenesis is mediated by neuroendocrine mechanisms, since infusion

of glucose directly into either the third ventricle or into the periphery produces similar

enhancement of metabolic rate (Le Feuvre et al., 1991), probably via activation of the

sympathetic nervous system whose sensitivity to glucose may play a role in the

development of obesity (Landsberg and Krieger, 1989; De Jonge and Bray, 2002).

1.7.3. Literature Pertinent to Glycosylated Haemoglobin

Historically, glycosylated haemoglobin (HbA1c) has been recommended

only for the determination of glucose control among persons who have already

received the diagnosis of diabetes. New clinical practice recommendations from the

American Diabetes Association (2010) advocate the use of glycosylated haemoglobin in

the diagnosis of diabetes, largely on the basis of the established association between

glycosylated haemoglobin and microvascular disease. Compared with fasting glucose,

glycosylated haemoglobin has several advantages as a diagnostic test; it has higher

repeatability (Phillipou and Phillips, 1993; Rohlfing et al., 2002; Selvin et al., 2007),

can be assessed in the non-fasting state and is the preferred test for monitoring glucose

control (American Diabetes Association, 2009). Long term prognostic data are also useful

for informing diagnostic cutoff points for asymptomatic conditions and there is evidence

that elevated glycated haemoglobin values may be a risk factor for macrovascular disease.

Glycosylated haemoglobin is being used with increasing frequency to monitor

long-term blood glucose control and its estimation provides an accurate index of the mean

concentration of blood glucose during the preceding two to three months

(Lapolla et al., 2005). Furthermore, haemoglobin has been considered as a model which

has provided insights into the non-enzymatic glycation of other tissue proteins

(Garlick et al., 1983). Other factors which influence the rate of glycosylation of proteins

include the prevailing concentrations of glucose and the half-life of the protein

(Lapolla et al., 2005).


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There is substantial interest in blood glucose concentrations because glucose reacts,

depending on blood glucose concentrations (Schnider and Kohn, 1980;

Brownlee et al., 1988), with amino groups of plasma and tissue proteins

(Amadori reaction) to form glycosylated proteins. These glycosylated proteins gradually

transform non-enzymatically into advanced glycosylation end products and have been

reported to result in altered protein function of the affected molecules. Glycosylation of

LDL, for example, was found to be associated with impaired receptor mediated uptake and

catabolism (Lyons, 1992) and glycosylation reactions may cause oxidative stress through

free radical generation (Giugliano et al., 1996; Jain and Palmer, 1997). Both diabetic micro

and macrovascular complications and increased atherosclerotic risk were reported to be

associated with advanced glycosylation end products (Schnider and Kohn, 1980;

Spagnoli et al., 1996).

Furthermore, glucose concentrations play an important role in the metabolic

syndrome. High serum glucose concentrations indicate the beginning of or existing

glucose intolerance and insulin resistance, which may result in type II diabetes.

The preclinical development of type II diabetes, however, is poorly understood and so far

there is little direct evidence that the same factors influencing metabolic control

in clinical diabetes might also affect the preclinical development of the disease

(Hannah and Howard, 1994). An increased risk of type II diabetes has been shown to be

associated with several dietary risk factors. High saturated fat intakes have been associated

with an increased risk of type II diabetes in various populations and diets high in complex

carbohydrates have been shown to protect against glucose intolerance and type II diabetes,

mainly because of their high fiber content (Virtanen and Aro, 1994).

Research on blood glucose concentrations was facilitated by the identification of

glycosylated haemoglobin (HbA1c) as a biomarker of long term glucose homeostasis

that reflects blood glucose concentrations over the previous 6 to 8 week

(Koenig and Cerami, 1980; Goldstein et al., 1982). In epidemiologic studies, this

biomarker has the advantage that a single assessment of HbA1c is suitable to

classify individuals according to their long term blood glucose concentrations

(Nathan et al., 1986).


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1.7.4. Literature Pertinent to Lipid Profiles

The lipid profile is a group of tests that are often ordered together to determine risk

of coronary heart disease. Lipids are used as hormones that play roles in regulating

metabolism of our body (Wardlaw, 1999). Lipid levels may be affected by diet, exercise,

smoking, certain medication e.g., beta blockers, thiazide diuretics, glucocorticoids and

concurrent disease states, e.g., kidney and liver diseases. They are tests that have been

shown to be good indicators of whether someone is likely to have a heart attack or stroke

caused by blockage of blood vessels or hardening of the arteries. Lipids are insoluble

(does not dissolve) in water but are soluble (dissolves) in alcohol and other solvents. When

dietary fats are digested and absorbed into the small intestine, they eventually reform into

triglycerides, which are then packaged into lipoproteins (Grundy, 1986; Sloop, 1999).

1.7.4.1. Effect on Cholesterol

Cholesterol is either obtained from the diet or synthesized in a variety of tissues,

including the liver, adrenal cortex, skin, intestine, testes and aorta. High dietary cholesterol

suppresses synthesis in the liver but not in other tissues (Linder, 1991; Salway, 1999).

1.7.4.2. Effect on Triglyceride

Carbohydrate is converted to triglyceride utilizing glycerol phosphate and acetyl

CoA obtained from glycolysis. Ketogenic amino acids, which are metabolized to acetyl

CoA, may be used for synthesis of triglycerides. The fatty acids cannot fully prevent

protein breakdown, because only the glycerol portion of the triglycerides can contribute to

gluconeogenesis (Murray et al., 1996).

Intramyocellular triglyceride (IMTG) in skeletal muscle has been implicated in

insulin sensitivity (Goodpaster et al., 1997; Jacob et al., 1999; He et al., 2001).

Indirect methods to measure IMTG by proton magnetic resonance spectroscopy (1H-MRS)

and by computed tomographic scanning have demonstrated that an excess of IMTG

is associated with insulin resistance (Perseghin et al., 1999; Virkamaki et al., 2001).

In accordance, studies that used more direct methods to estimate IMTG, i.e., by analyzing

skeletal muscle biopsies with oil red-O staining or by combining thin layer

chromatography and gas-liquid chromatographic analyses, have concluded that increased

IMTG is related to impaired insulin action (Pan et al., 1997; Levin et al., 2001 ).


http://www.labtestsonline.org/understanding/conditions/heart_attack.html

http://www.labtestsonline.org/understanding/conditions/stroke.html

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1.7.4.3. Effects on Phospholipids

The composition of structural lipids of the skeletal muscle cell membrane, i.e., the

phospholipids, may play a role for whole body insulin action (Borkman et al., 1993;

Helge and Dela, 2003; Haugaard et al., 2006). In vivo data from the rodent model and

in vitro data have suggested that changes in the phospholipid composition of cell

membranes are associated with the number of insulin receptors (Ginsberg et al., 1982),

the affinity of insulin to the insulin receptor (Grunfeld et al., 1981), membrane glucose

transport (Field et al., 1990) and the fact that a diet enriched with polyunsaturated fatty

acids (PUFAs) may increase long chain PUFAs (LCPUFAs) in muscle cell membrane and

decrease fasting plasma insulin (Storlien et al., 1991).

1.7.4.4. Effect on Free Fatty Acid

Fatty acids come from the diet, adipocytes (fat cells), carbohydrate and some

amino acids. After digestion, most of the fats are carried in the blood as chylomicrons.

The main pathways of lipid metabolism are lipolysis, β-oxidation, ketosis and lipogenesis.

Lipolysis (fat breakdown) and β-oxidation occurs in the mitochondria. It is a cyclical

process in which two carbons are removed from the fatty acid per cycle in the form of

acetyl CoA, which proceeds through the Krebs cycle to produce ATP, CO2 and water

(Salway, 1999; Wardlaw et al., 2002).

1.7.4.5. Effect on LDL-c, HDL-c and VLDL-c

The liver removes the chylomicron fragments, and the cholesterol is repackaged for

transport in the blood in very low density lipoproteins (VLDLs), which eventually turn

into low density lipoproteins (LDL). LDL cholesterol (LDL-c) is also called bad

cholesterol. Most LDL particles are absorbed from the bloodstream by receptor cells in the

liver. Cholesterol is then transported throughout the cells. Diets high in saturated fats and

cholesterol decrease the uptake of LDL particles by the liver. LDL particles are also

removed from the bloodstream by scavenger cells, or macrophages, which are white blood

cells that bury themselves in blood vessels such as arteries. Scavenger cells prevent

cholesterol from reentering the bloodstream, but they deposit the cholesterol in the inner

walls of blood vessels, eventually leading to the development of plaque

(Birtcher et al., 2000; Wardlaw et al., 2004).


http://www.faqs.org/nutrition/Diab-Em/Diet.html

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High density lipoproteins (HDLs) are a separate group of lipoproteins that contain

more protein and less cholesterol than LDL. HDL cholesterol is also called good

cholesterol. HDL is produced primarily in the liver and intestine and it travels in the

bloodstream, picks up cholesterol and gives the cholesterol to other lipoproteins for

transport back to the liver (Birtcher et al., 2000; Wardlaw et al., 2004).

1.7.4.6. Lipids and Coronary Heart Disease

Coronary heart disease is a global health problem. High intake of fats is risk factors

for its development (Keys, 1957). Excessive intake of fatty foods and lack of exercise lead

to obesity. Certain food items like fruits in association with exercise may be protective.

High plasma cholesterol is positively related to the risk of CHD (Gordon et al., 1977).

Elevation of plasma cholesterol is usually due to an increase in the level of LDL-c

(Grundy, 1987). Saturated fats and cholesterol in the diet play a major role in the causation

of hypercholesterolemia and act as a risk factor for CHD (Apgar et al., 1987).

Dyslipidemia is associated with hypertension, diabetes mellitus and obesity and is

one of the major risk factors for the development of cardiovascular disease. Obese people

tend to have relatively high triglyceride and low HDL-c. Obesity also raises the LDL-c

levels (Zicha et al., 1999; Franz et al., 2002; Martirosyan et al., 2007). Obesity and lack of

exercise tend to lead to insulin resistance. Insulin resistance has a negative effect on lipid

production, increasing VLDL-c, LDL-c and triglyceride levels in the bloodstream and

decreasing HDL-c. This can lead to fatty plaque deposits in the arteries enhancing the risks

for cardiovascular disease. Excess insulin increases renal sodium retention, which

increases blood pressure and can lead to hypertension (Khedmat et al., 2007).

1.8. LITERATURE PERTINENT TO ENZYMES

Enzymes make life on earth possible and all metabolic activities are under the

control of enzymes. The numerous chemical reactions, going on continuously in living

matter, would not be possible without enzymes, which are the tools of the living cells.

Moreover, a biological system with a balanced metabolic pattern is characterized by a

dynamic equilibrium between different enzymatic activities. Any abnormality in the

enzyme system and their coordination may lead to an inhibition or hyperfunction of the

organ concerned, which ultimately manifests as disease (Murray et al., 1996).


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Concentration of certain enzymes which are present almost in significant amount in

the plasma rises several time under certain pathological conditions thus the rise of

particular non-functional enzymes in blood may give an idea regarding the site of damage

or dysfunction of the tissues and thus help in the clinical diagnosis of toxicity.

Therefore, blood enzymes are asserted in diagnosis of the structural and functional status

of the body organs exposed to toxicants (Pant, 1999).

1.8.1. Literature Pertinent to Aspartate Aminotransferase and Alanine

Aminotransferase

An aspartate aminotransferase (AST) and alanine aminotransferase (ALT) test

measures the amount of this enzyme in the blood. AST and ALT are normally found in red

blood cells, liver, heart, muscle tissue, pancreas and kidneys. AST formerly was called

serum glutamic oxaloacetic transaminase (SGOT). Also, ALT was formerly called serum

glutamic pyruvic transaminase (SGPT). Low levels of AST and ALT are normally found

in the blood. When body tissue or an organ such as the heart or liver is diseased or

damaged, additional AST and ALT are released into the bloodstream. The amount of AST

and ALT in the blood is directly related to the extent of the tissue damage. After severe

damage, AST levels rise in 6 to 10 hours and remain high for about 4 days. Most increases

in ALT levels are caused by liver damage. The AST test may be done at the same time as a

test for alanine aminotransferase, or ALT. The ratio of AST to ALT sometimes can help

determine whether the liver or another organ has been damaged. Both ALT and AST levels

can test for liver damage (Pagana and Pagana, 2006; Chernecky and Berger, 2008).

Obesity is an important correlate of elevated serum ALT and AST levels

(Ruhl and Everhart, 2003), markers of liver injury in the general population. Emerging

evidences suggest a significant role of visceral adiposity and insulin resistance in inducing

fatty liver disease rather than overall adiposity (Kelley et al., 2003; Schaffler et al., 2005;

Iacobellis et al., 2007). High serum ALT and AST activity are widely used as reliable

surrogate markers of fatty liver (Kunde et al., 2005; Oh et al., 2006). Increased ALT

activity has been mainly associated with several components of the metabolic syndrome,

such as abdominal visceral obesity, impaired insulin sensitivity, raised fasting glucose and

unfavorable lipid pattern (Vozarova et al., 2002; Hanley et al., 2005).


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Importantly, several studies in adults have found that high levels of hepatic

enzymes, particularly ALT, predict later development of type II diabetes mellitus

(Vozarova et al., 2002). Recently Nadeau et al., (2005) reported a high prevalence of

elevated ALT among children with T2DM, unrelated to age, body mass index (BMI) and

glycemic index. Thus, elevations in ALT may be not only a marker of liver injury and a

surrogate for fatty liver disease but also an early indicator of looming diabetes.

As a biomarker of liver injury, ALT levels in a large multiethnic cohort of obese youth and

ALT elevations were related to alterations in insulin sensitivity, glucose tolerance and

inflammatory markers.

1.8.2. Literature Pertinent to Alkaline Phosphatase

Alkaline phosphatase (ALP) is a membrane bound enzyme found in a wide variety

of tissues, including liver, bone and the placenta and normally present in high

concentrations in growing bone and in bile. ALP is released into the blood during injury

and during such normal activities as bone growth and pregnancy. There are 4 ALP

isoenzymes in humans, each coded by a separate gene: tissue nonspecific (TNALP; also

known as liver-bone-kidney ALP), intestinal, placental and germ cell (Berger et al., 1987;

Millan, 1988). The enzyme is known to have phosphoprotein phosphatase and

transphosphorylation activity and might have an important role in bone mineralization

(Meyer-Sabellek et al., 1988). The serum levels of liver and bone ALP are used widely in

the diagnosis of hepato-biliary disease and various bone disorders

(Crofton, 1992; Price, 1993), respectively. It recently was reported that the TNALP

isoenzyme is present in human and murine pre-adipocytes and might have a role in the

intracellular accumulation of triglycerides that characterizes the process of adipogenesis

(Ali et al., 2005; 2006).

The existence of ALP in human pre-adipocytes is of interest because it is

conceivable that adipose tissue might be a source of serum ALP. Furthermore, the positive

relationship between measures of abdominal obesity and serum, liver enzyme levels

demonstrates that adipose tissue mass also can influence the release of liver products into

the circulation. Thus, the level of TNALP in serum might be influenced by total and

abdominal adipose tissue mass (Ali et al., 2006).


http://www.medterms.com/script/main/art.asp?articlekey=4918

http://www.medterms.com/script/main/art.asp?articlekey=33915

Introduction

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1.8.3. Literature Pertinent to Lactate Dehydrogenase

Lactate dehydrogenase (also called lactic acid dehydrogenase or LDH) is an

enzyme that catalyzes the conversion of lactate to pyruvate. This is an important step in

energy production in cells. Many different types of cells in the body contain this enzyme.

Some of the organs relatively rich in LDH are the heart, kidney, liver and muscle. As cells

die, their LDH is released and finds its way into the blood. Normal LDH levels vary with

age, being higher in childhood due to bone growth. It plays an important role in cellular

respiration, the process by which glucose (sugar) from food is converted into usable

energy for our cells (Borna et al., 2009).

LDH, a pyridine-linked enzyme found in virtually all animal and human tissues,

functions primarily in the metabolism of glucose, catalyzing the reduction of free pyruvate

to lactate during the last step of glycolysis, as well as the conversion of lactate to pyruvate

during gluconeogenesis. Its concentration is highest in liver followed in descending order

in skeletal muscle, heart and kidney (Puc et al., 1985). Malignant cells have a distinctive

type of metabolism in which the glycolytic sequence and the tricarboxylic acid cycle are

poorly integrated, hence the cells tends to utilize from five to ten times as much glucose as

do normal tissues, converting most of it into lactate (Lehninger, 2000). LDH exists in

many different cell systems and subsequent to tissue or cell damage, serum LDH levels

may be elevated.

1.9. LITERATURE PERTINENT TO ANTIOXIDANTS

Antioxidants are a type of complex compounds found in our diet that act as a

protective shield for our body against certain disastrous enemies (diseases) such as arterial

and cardiac diseases, arthritis, cataracts and also premature ageing along with several

chronic diseases. The recent researches on free radicals promise a revolutionary

improvement in health and life style of humans (Yoshikawa et al., 2000). Oxygen is

essential for aerobic life process. However, cell under aerobic condition are threatened

with the insult of reactive oxygen metabolites that are efficiently taken care by some

powerful agents in our human body (Ray and Husain, 2002). These agents, which lower

the burden of free radicals, are known as antioxidants.


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1.9.1. Types of Antioxidants

Rajkapoor et al. (2010) stated that basically these are classified into three

categories: Enzymatic and non-enzymatic antioxidants are found both in extra cellular as

well as intracellular environment. These are tactically arranged within the cell in order to

provide maximum protection against free radicals (superoxide dismutase, catalase,

glutathione peroxidase and glutathione reductase - enzymatic; glutathione and minerals -

non-enzymatic). Antioxidant derived from natural and dietary sources that aid in

antioxidant defense system, protecting plants against damage caused by active O2 formed

due to exposure to ultraviolet radiation. Certain seaweeds also function as antioxidants.

Our daily diet contains vegetables, fruits, tea, wine, etc., which possess compounds rich in

anti oxidative properties. Antioxidants from natural sources are chlorophyll derivatives,

essential oils, carotenoids, alkaloids, phytosterols, flavonoids, polyphenolics, tannins,

proanthocynidine, nitrogen containing compounds- alkaloids and indoles.

1.9.2. Oxidative Stress and Role of Antioxidants

Oxidation refers to transfer of electrons from a substance to an oxidizing agent.

Oxygen is an element indispensable for life. When cells use oxygen to generate energy,

free radicals are formed as a consequence of ATP production by the mitochondria.

These byproducts are generally called as reactive oxygen species (ROS) as well as reactive

nitrogen species (RNS) that result from the cellular redox process. At lower

concentrations, ROS and RNS exert beneficial effects as cellular response and immune

function. At high concentrations, they generate oxidative stress that can damage all cell

structures (Willcox et al., 2004; Halliwell, 2007). As oxidative stress is an important part

of many human diseases, the use of antioxidants in pharmacology is intensively studied,

particularly in the treatment of stroke and neurodegenerative diseases. Antioxidants are

widely used as ingredients in the dietary supplements in order to maintain health and to

prevent diseases such as cancer and coronary heart disease (Bielakovic et al., 2008).

1.9.3. Literature Pertinent to Lipid Peroxidation

Free radicals attacking biomembranes can lead to oxidative destruction of the

polyunsaturated fatty acids (PUFA) in the membranes by a process called lipid

peroxidation. Lipid peroxidation (LPO) involves the formation of lipid radical, oxidation

of unsaturated lipids and the eventual destruction of membrane lipids producing a variety

of break down products and deleterious effects (Metz, 1984).


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LPO is initiated by any chemical species that has sufficient energy to abstract

hydrogen atom of polyunsaturated fatty acids present in the phospholipid side chains.

Peroxidation of lipids exposed to oxygen is responsible not only for the deterioration of

foods, but also for damage to tissues in vivo (Nadkarni, 1992). Peroxidation of membrane

lipids is likely to lead to a disturbance of the membrane integrity (Richter, 1986;

Vliet and Bast, 1992).

LPO is a highly destructive process and alters the structure and function of cellular

membrane (Kale and Sitasawad, 1998). It is involved in a number of diseases and in

poisoning lipid peroxidation goes at a faster rate than normal ones. LPO, therefore, can be

used as a measure of oxidative damage. Peroxidation brings about changes in structure,

fluidity and permeability of membrane (Srivastavam et al., 1998) inactivates a number of

membrane bound enzymes and protein-receptors, swelling and alterations of respiratory

functions. Radiation induced apoptosis and lipid peroxidation are closely linked

(Agarwal and Kale, 2001).

Lipid peroxide mediated tissue damage has been observed in the development

of type II diabetes. It has been observed that insulin secretion is closely

associated with lipoxygenase derived peroxides (Metz, 1984; Walsh and Pek, 1984).

Increased concentration of thiobarbituric acid reactive species (TBARS) is also observed

in kidney during diabetes. Nakakimura and Mizuno (1980) have reported that the

concentration of lipid peroxides increases in the kidney of diabetic rats.

1.9.4. Literature Pertinent to Superoxide Dismutase

The first enzyme involved in the antioxidant defense is superoxide dismutase

(SOD). SODs are a family of metallo-enzymes. These are families of SOD; Cu-SOD,

Cu-Zn-SOD and Mn-SOD. SOD is an enzyme that disarms and destroys free radicals,

particularly superoxide. Cu and Zn are required in the functioning of cytosolic SOD and

manganese is required for the mitochondrial version. SOD mainly acts by quenching of

superoxide, an active oxygen radical, produced in different aerobic metabolism

(Li et al., 1995; Kizaki et al., 1993; MacMillan-Crow et al., 1998). O2 is the only substrate

for SOD. Each type of SOD has its own peculiarities; however all types of the enzyme

have similar properties (Oberley, 1984).


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SOD is considered to be a stress protein, which is synthesized in response to

oxidative stress. SOD can act as anti-carcinogens and inhibitor at initiation and promotion

stage in carcinogenesis. SOD is essential for the survival of aerobic cells. It catalytically

scavenges the superoxide radical and provides a defense against oxygen toxicity.

The cytosol of eukaryotic liver cells contains SODs with Cu2+

+ Zn2+

at the active site.

The mitochondrial SOD contains Mn3+

at the active site (Fridovich, 1995).

Superoxide is formed in the RBC by the auto oxidation of haemoglobin to

methemoglobin in other tissues; it is formed by the action of enzymes such as cytochrome

P450 reductase and xanthone oxidase. When stimulated by contact with bacteria,

neutrophils exhibit a respiratory burst and produce superoxide in a reaction catalyzed by

NADPH oxidase (Kalow and Grant, 1995). Hydrogen peroxide is subject to a number of

fats, the enzyme catalase present in many types of cells; convert it to H2O and O2.

Neutrophils possess a unique enzyme myeloperoxidase, which uses H2O2 and halides to

produce hypohalous acids (Katzung, 1998). Oxygen species are now thought to play an

important role in many types of cellular injury, some of which result in cell death.

Indirect evidence supporting a role for these species in generating cell injury is provided if

administration of an enzyme such as superoxide dismutase or catalase is found to protect

against cell injury in the situation under study (Nebert, 1997). SOD activity determination

will be utilized not only for the research of the mechanism that cause diseases but also for

the diagnosis and indication of health condition (Ukeda, 2003).

1.9.5. Literature Pertinent to Catalase

Catalase is a tetrameric haeme enzyme, which catalyses the decomposition of

highly poisonous H2O2 to water and oxygen protecting cells. It is found in almost all

animal cells except certain anaerobic bacteria (Voet and Voet, 1995). It is a haeme

containing enzyme and protects the cell from dangerous concentration of peroxide

produced by the respiratory chain and other oxidases. CAT contains four haem groups and

is found in blood, bone marrow, mucous membrane, peroxisomes in liver and kidney.

CAT

H2O2 + H2O2 2H2O + O2 (Triplet oxygen)

Relatively stable triple oxygen is formed in the enzymatic degradation of hydrogen

peroxide by catalase (Porter and Ingraham, 1974).


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Most of the in vitro studies suggested that this antioxidant functions as

promotion/transformation inhibitor in carcinogenesis. Catalase is found to reduce SCE

levels resulting from treatment with H2O2. CAT is found to act 104 times faster than

peroxidase. It is localized mainly in mitochondria and in sub cellular respiratory organelles

(Dixon and Webb, 1964). In liver, catalase is mainly localized in macrosomes and

microperoxisomes (Chance et al., 1979).

Increased rate of ROS production frequently elicits a response, an increase in the

level of antioxidants. Under high rate of free radicals input, the enzyme inactivation

prevails and enzymatic activities are reduced leading to autocatalysis of oxidative damage

process (Escobar et al., 1969).

1.9.6. Literature Pertinent to Glutathione

Glutathione is a tripetide (L-γ glutamyl-cystinyl glycine). Glutathione is the

predominant thiol compound in many cells, both prokaryotes and eukaryotes. Glutathione

exists in the reduced (GSH) and oxidized (GSSG) forms with GSH predominating inside

the cells. Glutathione participates in reactions that destroy hydrogen peroxide, organic

peroxide free radicals and certain foreign compounds. Glutathione participates by a

number of chemical mechanisms in the metabolism of various endogenous compounds.

It serves catalytically in some cases and as a reactant in others. Glutathione functions are

the transport of amino acids (Cornell and Meister, 1976).

A number of potentially toxic electrophilic xenobiotics (such as certain

carcinogens) are conjugated to nucleophilic GSH in reactions that can be represented as

follows:

R + GSH R-S-G

The enzymes catalyzing these reactions are called glutathione-S-tranferases and are

present in high amounts in liver cytosol and in lower amounts in other tissues. A variety of

glutathione-S-tranferases are present in human tissues (Ulusu et al., 2003). They exhibit

substrate specificities and can separate by electrophoretic and other techniques.

If the potentially toxic xenobiotics were not conjugated to GSH they would be free to

combine covalently with DNA and RNA. GSH is their important defense mechanism

against certain toxic compounds (Biemann, 1992).


Introduction

64 | P a g e

These enzymes have been shown to act as storage proteins and appear to be

responsible for the binding of bilirubin and its conjugates (Rana and Gupta, 1999).

It is generally agreed that the intra hepatic glutathione is able to afford protection against

liver dysfunction by atleast two ways, firstly as a substrate of glutathione peroxidase,

GSH serves to reduce a large variety of hydroperoxides before they attack unsaturated

lipids or convert already formed lipid hydroperoxides to the corresponding hydroxy

compounds. Secondly as a substrate of glutathione-S-transferase, it enables the liver to

detoxify many foreign compounds or their metabolites and to excrete the produce,

preferably into the bile (Arthur et al., 1987).

Glutathione (GSH) plays an important role in the maintenance of ascorbic acid in

reduced form and also acts as a cofactor for antioxidative enzymes (Kent, 1988).

Enhanced lipid peroxidation was followed by increased glutathione oxidation to form

oxidized glutathione (Ozols, 1990b). Glutathione peroxidases are selenoenzymes, which

catalyse the reduction of hydroperoxides at the expense of GSH (Ursini et al., 1995).

In this process, hydrogen peroxide is reduced to water whereas organic hydroperoxides are

reduced to alcohols. GPx not only decomposes hydrogen peroxides but is also capable of

interacting with lipid peroxidation (Koul and Kapil, 1994). In erythrocytes and other

tissues, the enzyme glutathione peroxidase, containing selenium as a prosthetic group,

catalyses the destruction of H2O2 and lipid hydroperoxides by reduced glutathione,

protecting membrane lipids and haemoglobin against oxidation by peroxides

(Stellwagen, 1990). The pentose phosphate pathway in the erythrocyte provide NADPH

for the reduction of oxidized glutathione to reduced glutathione reductase, a flavoprotein

enzyme containing FAD (Strickler et al., 1984).

1.10. LITERATURE PERTINENT TO LIVER

The liver, the largest organ of the human body, weighs approximately 1,500 g and

is located in the upper right corner of the abdomen. The major blood vessels, portal vein,

hepatic artery, lymphatics, nerves and hepatic bile duct communicate with the liver at a

common site, the hilus. From the hilus, they branch and re-branch within the liver to form

a system that travels together in a conduit structure, the portal canal. From this portal

canal, after numerous branching, the portal vein finally drains into the sinusoids, which is

the capillary system of the liver. Here, in the sinusoids, blood from the portal vein joins

with blood flow from end arterial branches of the hepatic artery. Once passed through the


Introduction

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sinusoids, blood enters the collecting branch of the central vein and finally leaves the liver

via the hepatic vein. The hexagonal structure with, in most cases, three portal canals in its

corners draining into one central vein, is defined as a lobule. The lobule largely consists of

hepatocytes (liver cells) which are arranged as interconnected plates, usually one or two

hepatocytes thick. The space between the plates forms the sinusoid. A more functional unit

of the liver forms the acinus. In the acinus, the portal canal forms the center and the central

veins the corners (Kakadiya, 2009).

Liver regulates various important metabolic functions. Hepatic damage is

associated with distortion of these metabolic functions (Wolf, 1999). Liver disease is still a

worldwide health problem. Unfortunately, conventional or synthetic drugs used in the

treatment of liver diseases are inadequate and sometimes can have serious side effects.

This is one of the reasons for many people in the world over including those in

developed countries turning to complementary and alternative medicine (CAM).

Many traditional remedies employ herbal drugs for the treatment of liver ailments

(Venkateswaran et al., 1997; Latha et al., 1999; Mitra et al., 2000).

1.10.1. Mechanisms of Hepatotoxicity:

Certain drugs will produce predictable liver damage in the majority of cases after

overdoses. In some cases the mechanism may involve the parent compound; in others a

metabolite may be responsible. Direct cytotoxicity is known to be the underlying cause of

liver damage in certain cases, whereas in others, immunological mechanisms or even a

mixture of both cytotoxicity and immunogenicity may be involved (Ingwale et al., 2009).

1.10.2. Hepatic Complications in Obesity

Liver disease complication is one of the most common causes of morbidity and

mortality in obesity patients. Liver plays an important role in normal glucose homeostasis

and a variety of liver conditions are associated with abnormal glucose homeostasis. This

association may explain the pathogenesis of the liver disease or of the abnormal glucose

homeostasis or may be purely coincidental (Kakadiya and Rathod, 2009).

1.10.3. Role of the Liver in Glucose Homeostasis

An appreciation of the role of the liver in the regulation of carbohydrate

homeostasis is essential to understand the many physical and biochemical alterations that

occur in the liver. The liver uses glucose as a fuel and also has the ability to store it as


Introduction

66 | P a g e

glycogen and synthesize it from non-carbohydrate precursors (gluconeogenesis). Glucose

absorbed from the intestinal tract is transported via the portal vein to the liver (Bjornstorp

and Sjostrom, 1978). Katz et al. (1983) suggested that most absorbed glucose is not taken

up by the liver but is rather metabolized via glycolysis in the peripheral tissues. In type II

diabetes, excessive hepatic glucose output contributes to the fasting hyperglycemia.

Increased gluconeogenesis is the predominant mechanism responsible for this increased

glucose output, while glycogenolysis has not been shown to be increased in patients with

type II diabetes (Consoli et al., 1989). Hyperglucagonemia has been shown to augment

increased rates of hepatic glucose output, probably through enhanced gluconeogenesis.

Insulin is metabolized by insulinase in the liver, kidney and placenta.

Insulin promotes glycogen synthesis (glycogenesis) in the liver and inhibits its breakdown

(glycogenolysis). It promotes protein, cholesterol and triglyceride synthesis and stimulates

formation of VLDL-c. It also inhibits hepatic gluconeogenesis, stimulates glycolysis and

inhibits ketogenesis. The liver is the primary target organ for glucagon action, where it

promotes glycogenolysis, gluconeogenesis and ketogenesis (Karem and Forsham, 1983).

1.11. LITERATURE PERTINENT TO KIDNEY The kidneys are important organs actively involved in excretory and regulatory

functions (Henegar et al., 2001). Kidneys are paired solid bean shaped organs; present on

each side of the retroperitoneal space, in mid position. Kidney is primarily responsible for

maintaining the stability of ECF volume, electrolyte composition and osmolarity

(Sherwood, 2007). The kidney is also responsible for the role of homeostasis,

excretion of waste product, maintaining the water balance, acid-base balance,

haemopoietic function, endocrine function, regulation of blood pressure and calcium level

(Sembulingam and Sembulingam, 2010). Kidney injury due to chemicals or infectious

agent may lead to glomerulonephritis, acute renal failure, chronic renal failure and

nephritic syndrome (Henegar et al., 2001).

Renal disease is a regular aspect of both insulin dependent (Type I) and noninsulin

dependent (Type II) diabetes mellitus (Ritz and Stefanski, 1996; Mozaffari et al., 1997) in

which the developed renal changes are attributed to a great extent to existing

hyperglycemia (Janssen et al., 1999; Usui et al., 2003). Progression of the disease process

results in end stage renal disease (ESRD) which accounts for approximately 35% of all

new admissions for renal replacement therapy (Ritz and Stefanski, 1996).


Introduction

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Acute renal failure (ARF) continues to be associated with significant morbidity and

mortality (Radhakrishnan and Kiryluk, 1997), renal ischemia/reperfusion (I/R) injury is a

major cause of ARF, which has to be faced in many clinical situations like renal

transplantation, partial nephrectomy, renal artery angioplasty, aortic aneurysm surgery and

elective urological operations that initiate a complex and interrelated sequence of events,

resulting in injury to, and the eventual death of renal cells (Thadhani et al., 1996;

Paller, 1998). Several factors have been implicated in the pathophysiological changes of

renal I/R injury including vascular/microvascular injury, endothelial dysfunction and

accelerated cell necrosis and granulocyte activation (Adam and Raij, 2000).

Control of blood glucose is crucial because of the proven link between HbA1c and

the rate of decline of the kidney function in diabetic nephropathy. Hyperglycemia caused

by the high carbohydrate diet necessitates the use of insulin; efforts to normalise the blood

glucose with insulin leads to increase of appetite and bodyweight (Henry et al., 1993);

the rise of bodyweight exposes the patient to the risk of obesity associated renal failure

(Hsu et al., 2006).

1.12. LITERATURE PERTINENT TO HEART

The heart is a myogenic muscular organ found in all animals with a circulatory

system that is responsible for pumping blood throughout the blood vessels by repeated and

rhythmic contractions. Heart failure (HF) is a condition in which a problem with the

structure or function of the heart impairs its ability to supply sufficient blood flow to meet

the bodies needs (McMurray and Pfeffer, 2005).

1.12.1. Obesity and Heart Failure

Cholesterol is a unique type of fat. The fat cells have a unique feature. They do not

multiply generally like other cells of the body until there is a shortage of these cells. Heart

supplies oxygen rich blood to the entire body. It has tube like structures called arteries and

veins. When the presence of cholesterol rises in the blood, the fat cells reach the heart and

make the surface area of the arteries and veins narrow. The organ works harder to pump

oxygen rich blood to the entire body and gets tired. This can lead to a fatal condition called

heart attack. Intake of low cholesterol food and regular exercise brings back the

efficiency of heart in pumping blood to normalcy. This reduces the risk of heart failure

(Kenchaiah et al., 2002).


http://en.wikipedia.org/wiki/Muscle

http://en.wikipedia.org/wiki/Organ_(anatomy)

http://en.wikipedia.org/wiki/Animal

http://en.wikipedia.org/wiki/Circulatory_system

http://en.wikipedia.org/wiki/Circulatory_system

http://en.wikipedia.org/wiki/Blood

http://en.wikipedia.org/wiki/Blood_vessel

Introduction

68 | P a g e

1.12.2. Obesity and Coronary Heart Disease

Until recently the relation between obesity and coronary heart disease was viewed

as indirect (Lew and Garfinkel, 1979), both obesity and coronary heart disease risk

including hypertension; dyslipidemia, particularly reductions in HDL-c

and impaired

glucose tolerance or noninsulin dependent

diabetes mellitus. Insulin resistance and

accompanying hyperinsulinemia

are typically associated with these co-morbidities

(Reaven, 1988). Although most

of the co-morbidities relating obesity to coronary

artery disease increase as BMI increases, they also relate to body fat distribution

(Manson et al., 1995).

1.12.3. Lipid Metabolism and Heart Failure

Under physiologic conditions, most triglycerides are stored in adipocytes with only

minimal accumulation of lipids in other tissues such as the liver or muscle.

Increased stores of triglycerides are detectable in the myocardium of animals with obesity

and diabetes (Zhou et al., 2000). This finding has been reproduced in patients with heart

failure and diabetes (Sharma et al., 2004) and correlates with the degree of obesity

(Peterson et al., 2004; McGavock et al., 2007).

1.13. SCOPE OF THE PRESENT INVESTIGATION

With the exception of one or two, most of the conventional anti-obesity drugs

produce some sort of side effect. Plants have been the sources of drugs in Indian systems

of medicine and of other ancient system in the world. Medicinal plants have the advantage

of having little or no side effects. Some of them are being used in traditional system of

medicine for hundreds of years in many countries of the world. Hence, the herbal

medicines geriforte, haritaki and ayurslim were selected to observe their effect on obesity

induced changes in metabolic hormones, biochemical parameters, oxidative stress and

antioxidant profiles.

Considerable amount of drug screening studies are being undertaken on various

organ systems. Of these, liver and kidney seem to be the preferred organs that have been

studied due to presence of complex enzyme system which play a major role in metabolism,

detoxification, glycogen storage, plasma protein synthesis etc. Heart is the most affected

organ due to obesity. So these three organ systems become the model of these drug

actions.


Introduction

69 | P a g e

1.13.1. Main Aims of the Study are ascertain that

The obesity induction raises the body weight, organ weight (liver, kidney and heart)

as well as fat pad weights (abdominal and perirenal).

The herbals, geriforte, haritaki and their combination as well as ayurslim reduce the

body weight, organ weight and fat pad weights.

The obesity alters the hormones of metabolism like insulin, adrenal glucocorticoids

and also the pituitary-thyroid axis.

The herbal treatments can restore the hormonal levels to normalcy setting right the

altered metabolism as well as the metabolic hormones.

The obesity affects the haematological parameters like RBC, WBC, Hb and MCH.

The herbal treatment alleviates the obesity induced toxicity by restoring the above

haematological parameters.

The obesity induction alters the serum biochemical parameters like total protein,

glucose, glycosylated haemoglobin, total cholesterol, triglycerides, phospholipids, free

fatty acids, HDL-c, LDL-c, VLDL-c, aspartate amino transferase, alanine amino

transferase, alkaline phosphatase and lactate dehydrogenase.

The herbal treatments restored the above said biochemical parameters to near

normalcy.

The obesity affects liver, kidney and heart by the way of altering the lipid profiles in

them.

The experimental drugs restructure the obesity induced changes in lipid profile.

The obesity affects the serum LPO and antioxidant parameters like SOD, CAT and

GSH.

The herbal treatment alleviates the obesity induced toxicity by restoring the above

antioxidant parameters.


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70 | P a g e

The obesity adversely affects the liver and kidney by enzymes like AST, ALT, ALP

and LDH, the generation of oxidative stress due to changes in lipid peroxidation and

the representative enzymes (superoxide dismutase, catalase and glutathione) involved

that reveal the extent of toxicity if any.

The herbals improve the obesity induced toxicity by restoring the above liver

enzymological and kidney antioxidant parameters in the major organ system by their

enzyme and antioxidant properties.

The obesity induces structural changes in the liver, kidney and heart.

The herbals restore the histoarchitecture of the above tissues.


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