cruel capsule - virgo · pdf filepharmacological evaluation of cruel capsule on rat mammary...

Pharmacological evaluation

of

CRUEL CAPSULE

on Rat Mammary Gland Cancer

Sponsor: VIRGO UAP PHARMA PVT. LTD. Survey No. 423/98-B, Mahagujarat Ind. Estate, Sarkhej-Bavala Highway., Vill.:Moraiya, Tal.: Sanand, Dist.: Ahmedabad-382213 ( Gujarat ), INDIA

Study Center: Department of Pharmacology, Institute of Pharmacy, Nirma University. S.G. Highway, Ahmedabad-382481 ( Gujarat ), INDIA

Study Report

Pharmacological evaluation

of

CRUEL CAPSULE on Rat Mammary Gland Cancer

Study Director

Prof. Manjunath D. Ghate

Principal Investigator

Dr. Snehal S. Patel ………………………………………

Department of Pharmacology, Institute of Pharmacy,

Nirma University, Ahmedabad-382481,

Gujarat ( INDIA )

LIST OF CONTENTS

SR. NO TITLE

PAGE NO.

1 SUMMARY

2 INTRODUCTION

3 MATERIALS AND METHODS

4 RESULTS

5 DISCUSSION AND CONCLUSION

3

Summary Breast cancer is the most frequently diagnosed cancer and the leading cause of cancer death in

females worldwide. The present study was designed to evaluate the effect of oral treatment with

herbomineral formulation on DMBA-induced mammary carcinogenesis in rat. 50 days

postpartum female Sprague Dawley rats grouped into control, control treated with CRUEL

CAPSULE (100mg/kg/day, p.o.), disease control, disease treated with CRUEL CAPSULE

(100mg/kg/day, p.o.). Breast cancer was induced by oral administration of 60 mg/kg

dimethylbenz[a]anthracene (DMBA). Animals were palpated twice a week starting five weeks

after DMBA administration in order to record the presence, location, size, and date of detection

for all tumors and treatment was carried out through out the experiment. After 48 hours of the

last dose, blood was collected and serum was separated for estimations of Cancer Antigen 15.3

(CA 15.3), Carcino Embryonic Antigen (CEA), Creatine kinase (CK), lactate dehydrogenase

(LDH), Gamma glutamyl transferase (GGT), C-Reactive Protein (CRP), Erythrocyte

Sedimentation Rate (ESR), % Haemoglobin. Animals were sacrificed when the tumor diameter

reached one inch, animals became moribund or rats reached 18 weeks post-DMBA treatment.

Tumor was isolated for histopathological examinations and antioxident studies. Administration

of DMBA caused significant increase in cancer biochemical markers LDH and GGT. Treatment

showed significant decrease in LDH and GGT as compared to disease control group. The

treatment also showed significant decrease in MDA and significant increase in GSH and SOD

levels in breast homogenate as compared to disease control group. Treatment also caused a

significant decrease in inflammatory markers like, CRP, ESR and decreases hemoglobin levels

as compared to disease control group. Histopathological examinations shows hyperplastic and

well demarcated tubular adenoma which was inproved by treatment with CRUEL CAPSULE.

From the present investigations, we can conclude that the CRUEL CAPSULE inhibits the tumor

progression in animal model of breast cancer. Therefore the use of CRUEL CAPSULE as a

neo-adjuvant or adjuvant therapy may be beneficial.

Dr. Snehal Patel Principal investigator, Department of Pharmacology, Institute of Pharmacy, Nirma University

Prof. Manjunath Ghate, Study Director, Institute of Pharmacy, Nirma University

4

Introduction: Although the second leading cause of cancer mortality in women is breast cancer. Extensive

investigation has identified breast cancer susceptibility genes that can be inherited, but these

appear to play a minor role in most cases of breast cancer. The geographic variation in incidence

of breast cancer suggests that environmental and dietary factors play a significant role. A major

class of environmental carcinogens is a family of structurally related chemicals, the polycyclic

aromatic hydrocarbons (PAH). These carcinogens and related halogenated compounds have been

implicated in mammary tumorigenesis by epidemiological and laboratory studies. DMBA (7,12-

dimethylbenz [a] anthracene) is a prototypical PAH that has been used to promote tumors in

laboratory animals. Mammary tumors can be produced in rodents following administration of

DMBA by oral gavage, leading to up-regulation of the cellular cytosolic receptor for DMBA, the

aryl hydrocarbon receptor (AhR). Recent surveys of chemoprevention documented a variety of

phytochemicals that displayed activity in animals and may be candidates for chemopreventive

studies in humans. The 7,12-dimethylbenz[a]anthracene (DMBA)-induced mammary cancer

models in rats have been employed to identify substances with chemopreventive activity in many

herbal drugs and formulations. The current focus of chemopreventive research is on

intermediate biomarkers capable of detecting early changes that can be correlated with inhibition

of carcinogenic progression. The assessment of ROS-induced lipid peroxidation and the status of

antioxidants are widely used in the detection and evaluation not only of environmental

carcinogens, but also of presumptive antimutagens and anticarcinogens. The present study was

designed to evaluate the effect of oral pretreatment with the herbomenaral formulation in animal

model of mamery gland cancer. This study was carried out to lend scientific evidence to the

efficacious claim of the use of Ayurvedic propritory medicine in the management of breast

cancer in folklore medicine.

Materials and methods: Experimental Animals:

Seven- to eight weeks old female SD rats weighing 170-200 g were obtained from the animal

facility of Zydus research centre, Ahmedabad, were housed in a pathogen-free environment at

the animal house of Institute of pharmacy, Nirma University. Animals were maintained under

well controlled temperature 22 ± 8oC & humidity 55 ± 5% and 12h/12h light/dark cycle. They

kept in well ventilated animal house under natural photoperiodic condition in polypropylene

cages with free access to food and water ad libitum. All experiments and protocols described in

5

present study were approved by the Institutional Animal Ethics Committee (IAEC) of Institute

Of Pharmacy, Nirma University, Ahmedabad as per guidelines of Committee for the Purpose of

Control and Supervision of Experiments on Animals (CPCSEA), Ministry of Social Justice and

Empowerment, Government of India. All the experiments described in present study were

conducted as per protocol number IPS/PCOL/CONS11-12/1003 dated 10-08-2011.

Experimental protocol:

At 50 days postpartum, 32 female SD rats grouped into control, control treated with CRUEL

CAPSULE (30mg/kg/day, p.o.), disease control, Disease treated with CRUEL CAPSULE

(30mg/kg/day, p.o.). Breast cancer was induced by oral administration of 60 mg/kg

dimethylbenz[a]anthracene (DMBA) (Timothy et al., 2006), a dose sufficient to cause 100%

tumor incidence in the disease control group and disease treated group over the course of the

study. The DMBA was dissolved in sesame oil at a stock solution of 60 mg/ml. Animals were

palpated twice a week starting five weeks after DMBA administration in order to record the

presence, location, size, and date of detection for all tumors. Animals were sacrificed when the

tumor diameter reached one inch, animals became moribund, or rats reached 18 weeks post-

DMBA treatment. Sections from the tumors were fixed in 10 % neutral buffered formalin and

embedded in paraffin. Tumor blocks were sectioned and fixed on slides to be evaluated

histopathologically.

Results : Effect of CRUEL CAPSULE on body weight :

In the normal control group change in body weight was found to be 65.00±7.63. A significant

decrease in body weight was found to be in disease control group as compared to normal control

group. Whereas the body weight was found to be increased in control treated with CRUEL

CAPSULE and disease treated with CRUEL CAPSULE (Table 1, Figure 1)

Table 1. Effect of CRUEL CAPSULE on body weight of SD rats

Group Change in Body weight gain

NC 65.00 ± 7.63

DC -52.50 ± 4.78*

FORM 46.0 ± 5.09

D+FORM 54.29 ± 10.43#

6

Values expressed as Mean±SEM of 6 animals each, * Significantly different from normal control

(p<0.05), # Significantly different from disease control group (p<0.05). NC-Normal control, DC-

Disease control, FORM- Normal animals treated with CRUEL CAPSULE (30mg/kg),

D+FORM- Diseased animal treated with CRUEL CAPSULE.

Figure 1. Effect of CRUEL CAPSULE on body weight



Disease control, FORM-Normal animals treated with CRUEL CAPSULE (30mg/kg), D+FORM-

Diseased animal treated with CRUEL CAPSULE.

Effect of CRUEL CAPSULE on biochemical parameters in serum:

1. Lactate Dehydrogenase (LDH):

In the normal control group, LDH levels were found to be 821.8 ± 38.36 U/L. A significant

increase in the LDH level was found in the disease control group as compared to normal control

group. Whereas in the disease treated with CRUEL CAPSULE group LDH level was found to be

significantly reduced as compared to disease control group (Table 2, Figure 2).

Table 2 Effect of CRUEL CAPSULE on Lactate Dehydrogenase level.

Group LDH level (U/L)

NC 821.8 ± 38.36

DC 1479± 127.5*

FORM 966.4±88.11

D+FORM 762.7 ± 116.6#

7



Disease control, FORM-Normal animals treated with CRUEL CAPSULE (30mg/kg), D+FORM-

Diseased animal treated with CRUEL CAPSULE.

Serum LDH Levels

NC DCFORM

D+FORM

0

500

1000

1500

2000

LDH

(U/L

)

Figure 2. Effect of CRUEL CAPSULE on Lactate Dehydrogenase level





2. Gamma Glutamyl Transferase (GGT) Activity:

In the normal control group, GGT activity was found to be 3.96 ± 0.26 U/L. A significant

increase in the GGT activity was found in the disease control group as compared to normal

control group. Whereas in the disease treated with CRUEL CAPSULE group GGT activity was

found to be significantly reduced when compared to disease control group (Table 3, Figure 3).

8

Table 3. Effect of CRUEL CAPSULE on serum Gamma Glutamyl Transferase activity

Group GGT activity (U/L)

NC 3.96 ± 0.26

DC 6.92 ± 0.96*

FORM 4.191± 0.36

D+FORM 4.64 ± 0.35#




D+FORM- Diseased animal treated with CRUEL CAPSULE

Serum GGT Activity

NC DCFORM

D+FORM

0

2

4

6

8

10

*

#

GG

T U

/L

Figure 3 Effect of CRUEL CAPSULE on serum Gamma Glutamyl Transferase activity





9

Effect of CRUEL CAPSULE on oxidative stress parameters in mammary gland

homogenate:

1. Lipid peroxidation/Malondialdehyde (MDA) Level:

In the normal control group, MDA level was found to be 1.41 ± 0.07 nmol/mg. A significant

increase in the MDA level was found in the disease control group as compared to normal control

group. Whereas in the disease treated with CRUEL CAPSULE group, MDA level was found to

be significantly reduced as compared to disease control group (Table .4, Figure 4).

Table 4. Effect of CRUEL CAPSULE on MDA levels

Group MDA level (nmol/mg protein)

NC 1.41 ± 0.07

DC 2.56 ± 0.09*

FORM 1.51 ± 0.07

D+FORM 2.00 ± 0.15#





MDA Levels

NC DCFO

RM

D+FORM

0

1

2

3*

#

Aver

age

MD

A (n

mol

es/m

g pr

otei

n)

Figure 4. Effect of CRUEL CAPSULE on Malondialdehyde levels

10





2. Reduced Glutathione (GSH)

In the normal control group, GSH level was found to be 7.28 ± 0.69 µg/mg. A significant

decrease in the GSH level was found in the disease control as compared to normal control group.

Whereas in the disease treated with CRUEL CAPSULE group, GSH level was found to be

significantly increased as compared to disease control group (Table 5, Figure 5).

Table 5. Effect of CRUEL CAPSULE on Reduced Glutathione levels

Group GSH (µg/mg protein)

NC 7.28 ± 0.69

DC 2.68 ± 0.46*

FORM 6.52 ± 0.70

D+FORM 5.10 ± 0.28#





GSH LEVELS

NC DC

FORM

D+FORM

0

2

4

6

8

10

*

#

Aver

age

Glu

tath

ione

con

c. (µ

g/m

g pr

otei

n)

Figure 5. Effect of CRUEL CAPSULE on Reduced Glutathione Levels



11



3. Super oxide dismutase (SOD):

In the normal control group, SOD level was found to be 2.49 ± 0.16 U/mg. A significant

decrease in the SOD level was found in the disease control group as compared to normal control

group. Whereas in the disease treated with CRUEL CAPSULE group, SOD level was found to

be significantly increased as compared to disease control group (Table 6, Figure 6).

Table 6. Effect of CRUEL CAPSULE on Super oxide dismutase levels

Group SOD (U/mg protein)

NC 2.49 ± 0.16

DC 1.61 ± 0.10*

FORM 2.55 ± 0.07

D+FORM 2.32 ± 0.06#





SOD LEVELS

NC DC

FORM

D+FORM

0

1

2

3

*

#

SOD U

nits

/mg

Prot

ein

Figure 6. Effect of CRUEL CAPSULE on tissue Super oxide dismutase levels





Effect of CRUEL CAPSULE on serum biochemical Markers:

12

1. Carcino Embryonic Antigen (CEA) level:

In the normal control group, carcino embryonic antigen (CEA) level was found to be below 0.5

ng/ml which is not a detectable level. In disease control group CEA level was found to be 0.5

ng/ml.

2. Cancer Antigen 15.3 levels:

In all the groups CA 15.3 levels were found to be below 0.5 ng/ml which is not a detectable

level.

Effect of CRUEL CAPSULE on inflammatory markers

1. C-Reactive Protein Level:

In the normal control group, CRP level was found to be 199.2± 20.94 mg/l. A significant

increase in the CRP level was found in the disease control group as compared to normal control

group. Whereas in the disease treated with CRUEL CAPSULE group CRP level was found to be

significantly reduced as compared to disease control group (Table 7, Figure 7).

Table 7. Effect of CRUEL CAPSULE on C -Reactive Protein Levels

Group CRP level (mg/l)

NC 199.2± 20.94

DC 300.9 ± 10.22*

FORM 218.3 ± 5.90

D+FORM 219.5± 9.02#





13

CRP LEVEL

NC DCFO

RM

D+FORM

0

100

200

300

400

*#

CR

P (m

g/l)

Figure 7. Effect of CRUEL CAPSULE on C -Reactive Protein levels





2. Erythrocyte Sedimentation Rate (ESR):

In the normal control group, ESR was found to be 2.24 ± 0.06 mm/hr. A significant increase in

the ESR was found in the disease control group as compared to normal control group. Whereas

in the disease treated with CRUEL CAPSULE group ESR was found to be significantly reduced

as compared to disease control group (Table 8, Figure 8).

Table 8. Effect of CRUEL CAPSULE on Erythrocyte Sedimentation Rate

Group ESR mm/hr

NC 2.24 ± 0.06

DC 3.47 ± 0.27*

FORM 2.42 ± 0.05

D+FORM 2.51 ± 0.13#





14

Erythrocyte Sedimentation Rate

NC DCFO

RM

D+FORM

0

1

2

3

4 *

#

ESR

(mm

/Hr)

Figure 8 Effect of CRUEL CAPSULE on Erythrocyte Sedimentation Rate





Tumor assessment:

15

Tumor of Disease control group Tumor of CRUEL CAPSULE treated group

Figure 9 Rat bearing mammary gland tumour

Histopathological Evaluation:

A B

C D

Figure 10: A, B, C, D- Histo architectute of mammary gland of Normal control, Disease

control, Disease treated with CRUEL CAPSULE and Control treated with CRUEL

CAPSULE respectively.

16

A. Mammary Gland of Normal control:

Normal ductular and alveolar structure of female mammmary gland. No sign of inflammation or

tissue damage, erosions / hyperplasic lesions was observed.

B. Mammary Gland of Disease control:

Mammary gland adenoma: Showing hyperplastic and well demarcated tubular adenoma. The

tubular and alveolar epithelium shows diffuse hyperplasia with minimal rim of scletan muscle.

Normal cellular detils preserved along with no metaplasic lesions was observed.

C. Mammary Gland of Disease treated with CRUEL CAPSULE:

Mild to moderate ductular and alveolar structure of female mammmary gland. No sign of

inflammation or tissue damage, erosions / hyperplasic / neoplastic lesions was observed.

D. Mammary Gland of Normal control treated with CRUEL CAPSULE:

Normal ductular and alveolar structure of female mammmary gland. No sign of inflammation or

tissue damage, erosions / hyperplasic / neoplastic lesions was observed.

Discussion: Human breast cancers represent a heterogeneous group of tumors that are diverse in behavior,

outcome and response to therapy (Rakha et al., 2007). Breast cancers defined as tumors having

the expression of estrogen receptors, progesterone receptors, HER-2, and the basal-like type

(Dent et al., 2007). Breast cancer tends to occur in younger women and BRCA1 carriers.

Clinically, they behave more aggressively, have a poor prognosis (Dolle et al., 2009) and have

an increased risk of distant recurrence (visceral and brain metastases) (Rakha et al., 2007).

Present chemotherapeutics do not specifically target tumor cells, but interfere with cell division

or inhibit enzymes involved in DNA replication or metabolism of normal dividing cells of

rapidly regenerating tissues, such as those in bone marrow, gut mucosa and hair follicles. This

limits the therapy of breast cancer with present chemotherapeutics. Considering the side effects

associated with synthetic chemotherapeutics and the potential of traditional and folk medicines in

the management of breast cancer. There is an increasing need to develop the treatment strategy

for breast cancer.

The word ‘cancer’ may be new to the 5,000 year old Indian system of medicine, Ayurveda,

which relies on natural substances for healing. But ancient Ayurvedic classics are aware of the

clinical features, resembling cancer, with the names such as Apachi, Gulma, Granthi, and

Arbuda. The excruciating experience of dying cancer patients can be ameliorated by making use

of Ayurvedic principles. Ayurveda can be helpful in the management of cancer in many ways

17

such as—prophylactic, palliative, curative and supportive. Ayurvedic medicines help to improve

the quality of life of the patients

The present study was carried out to lend scientific evidence to the efficacious claim of the use

of Ayurvedic propritory medicine in the management of breast cancer.

CRUEL CAPSULE contains seven medicinal plants (Piper nigrum, Boerhavia diffusa,

Glycyrrhiza glabra, Adhatoda vasica, Syzygium aromaticum, Moringa pterygosperma,

Tinospora cordifolia) each having proved anticancer activities. According to various preclinical

and clinical studies, such plants have shown very impressive effects on protecting body from the

side effects (of chemo and radio therapies) like alopecia, nausea, constipation, anorexia and

vomiting. The patients who receive Ayurvedic treatment along with chemotherapy have shown

more regularity in maintaining the chemotherapy cycles as the hematological levels can be

maintained up to a normal state. Several research studies on cancer carried out have shown that

Ayurvedic preparations are very effective as chemo-preventive and radio-preventive agents.

The formulation also contains minerals like Sarveswar Parpati, Suvarna Bhasma, Rasa Kapoor,

Abhrak Bhasma, Hira Bhasma, Rasasindur, Tamra Bhasma, Panna Bhasma. The active

principles of Sarveswar Parpati, Rasa Kapoor, Abhrak Bhasma, Rasasindur strengthens the

tissue, heals the ulcers, decreaces tumer growth by inhibition of angiogenesis, improves the

general condition and reduces the weakness of the carcinoma. Thus they reduces side effects

produced during treatment. The CRUEL CAPSULE also contains Hirak Bhasma, Suvarna and

Panna Bhasma, which is a very famous Ayurvedic medicine used in three doshas or imbalances

related to vaata, pitta and kapha. all these Bhasmas which are used in CRUEL CAPSULE have

the ability to reduce the three doshas or imbalances related to vaata, pitta and kapha.

It has been previously reported that medicinal plant present in CRUEL CAPSULE suppresses

the growth of experimental tumors in variety of tumor models in rodents and cell lines. However,

In the present investigation we found benificial effect of CRUEL CAPSULE on mammary

tumour in female Sprague Dawley rats induced by DMBA.

DMBA causes decrease in body weight of SD rats as compared to normal control group, These

results are in consistent with previous reports with DMBA on SD rats (Davis et al., 2001). A

yellow color liquid was found in the intestine of the disease control group when dissected, so it

was suggested that DMBA causes severe GI toxicity in the given dose of 60 mg/kg. Treatment

with CRUEL CAPSULE produced an improvement in body weight indicates that it reduces

toxicities produced by DMBA.

18

Lactate Dehydrogenase (LDH) is an enzyme that, under anaerobic conditions, catalyzes the

reversible transformation of pyruvate to lactate. Upregulation of LDH ensures an efficient

anaerobic/glycolytic metabolism for tumor cells and reduced dependence on oxygen (Rees et al.,

1960). Here increased LDH level in the disease control group may be due to increased hypoxic

stress and anaerobic metabolism for cancer cells, which ultimately cause the release of Hypoxia

inducible factor (HIF) and formation of new blood vessels for the growth of developing tumour.

Decreased LDH level observed in CRUEL CAPSULE treated group suggests the reduction in

hypoxic tress and anaerobic metabolism. Moreover LDH plays an important role in inflammation

and DNA damage. In this sense decreased LDH in treated groups plays beneficial role in the

prevention of inflammation and ultimately mammary gland cancer.

Gamma-glutamyl transferase (GGT) is a cellular enzyme with wide tissue distribution in the

body, primarily in the breast, kidney, pancreas, liver and prostate. Measurements of gamma-

glutamyl transferase (GGT) activity are used in the diagnosis and treatment of metastatic

tumors. Elevated activities of GGT observed in the DMBA-treated group are indicative of

DMBA-induced damage and subsequent leakage of these enzymes into circulation (Anbuselvam

et al., 2007). GGT act as useful markers for hepatic metastasis from breast and colon primaries.

Treatment with CRUEL CAPSULE to DMBA-treated animals significantly decreased GGT

activities indicating a significant cytoprotective effect on the epithelial cells of mammary gland.

C-reactive protein (CRP) is a general marker for inflammation. It is synthesized in hepatocytes

and belongs to the family of acute-phase proteins, the concentration of which change in response

to neoplasia. These changes are up-regulated by cytokines such as interleukin-6 (IL-6),

interleukin-8 (IL-8) and tumour necrosis factor (TNF) (Jones et al., 2007). In our study elevated

CRP level was found in serum of disease control group. Whereas CRP level was found to be

reduced in CRUEL CAPSULE treated groups. So it was suggested that CRUEL CAPSULE

interfere with one of the pathways of synthesis of IL-6 or IL-8 or TNF by inhibition of release of

biomarkers and thereby neoplasia.

ESR is used to detect inflammation in the body. Higher sedimentation rates indicate the presence

of inflammation (Guo et al., 2002). In our study high ESR was observed in disease control

group. Whereas ESR was found to be reduced in disease treated with CRUEL CAPSULE.

Therefore it was suggested that CRUEL CAPSULE reduces the inflammation there by reduces

the progression of mammary gland cancer by inhibition of progress of the disease.

19

Reactive oxygen species generation is a major factor involved in all steps of carcinogenesis, i.e.

initiation, promotion and progression (Trush et al., 1991). Oxidant-antioxidant balance impacts

the rate of cell proliferation and tumor cells generally display low levels of lipid peroxidation

which in turn can stimulate cell division and promote tumor growth (Rice et al., 1993). Oxidative

stress induced due to the generation of free radicals and/or decreased antioxidant level in the

target cell and tissues has been suggested to play an important role in carcinogenesis.

while targeting the fast-multiplying mutant cells, the toxicity they cause to normal tissues of the

body proves as an obstacle. The use of antioxidants during cancer treatment enhances therapy by

reducing the generation of free radicals. Natural drugs, which are used in Ayurveda, have also

been proved to have antioxidant properties.

Free radicals are involved both in the initiation as well as promotion stage of tumorigenesis and

their biochemical reactions in each stage of the metabolic process are associated with cancer

development (Huang et al., 1999). It is evident from the results that increased level of MDA was

found in cancer bearing animals when compared to control group. On the contrary, reduced level

of MDA was observed in disease treated with CRUEL CAPSULE indicating that it is a potent

free radical scavenger. During the process of neutralizing toxic reactants, CRUEL CAPSULE

protects proteins, lipids, mitochondrial DNA and nuclear DNA from oxidative damage.

Antioxidants act as the primary line of defense against ROS and suggest their usefulness in

eliminating the risk of oxidative damage induced during carcinogenesis. The medicinal plants

present in CRUEL CAPSULE Piper nigrum, Boerhavia diffusa, Glycyrrhiza glabra, Adhatoda

vasica, Syzygium aromaticum, Moringa pterygosperma, Tinospora cordifolia reported to have

proved antioxidant activities. SOD and CAT acts as mutually supportive antioxidative enzymes,

which provide protective defense against reactive oxygen species. The present study reveals that

the activity of SOD is depleted in the cancer-bearing animals, which may be due to altered

antioxidant status caused by carcinogenesis (Van et al., 1997).

The non-enzymatic antioxidant systems are the second line of defense against free radical

damage. GSH and GSH depended enzymes are involved in scavenging the electrophilc moieties

involve in the cancer initiation. GSH acts directly as a free radical scavenger by donating a

hydrogen atom and thereby neutralizing the hydroxyl radical. It also reduces peroxides and

maintains protein thiols in the reduced state (sies, 1996). GSH serves as a marker for evaluation

of oxidative stress and it act as an antioxidant at both intracellular and extra cellular levels. We

also observed decreased activities of GSH in cancer-bearing animals. CRUEL CAPSULE

20

showed increased level of the GSH , which clearly suggest their antioxidant property and there

by anticancer activity.

Tumor cells of mammary gland secrets tumor markers in serum mainly Carcino Embryonic

Antigen (CEA), Cancer Antigen 15.3 (CA 15.3). CEA is one of the most widely used tumor

marker in breast cancer. It is also a specific cancer marker which shows the state and

progression of malignancy.

CA 15.3 is a carbohydrate antigen present on the surface or secreted by the carcinoma cells. This

test is used as a valuable tumor marker for already diagnosed breast cancer (Beedu et al., 2005).

In our study in all the groups CA 15.3 levels were found to be below 0.5 ng/ml which is not a

detectable level. This observed low level of CA 15.3 might be due to its appearance in advanced

stage of malignancy.

CEA is a glycoprotein present in foetal gut & adenocarcinoma. Plasma CEA levels increase with

increasing age and with smoking. It can also used to detect carcinoma metastasis to breast.

Values 5-10 times the upper limit in patients with symptoms should be considered suggestive of

the presence of cancer. Post operative monitoring of plasma CEA levels has proved valuable for

detection of recurrence (Beedu et al., 2005). In our study in the normal control group, carcino

embryonic antigen (CEA) level was found to be below 0.5 ng/ml which is not a detectable level.

In disease control group CEA level was found to be 0.5 ng/ml. This observed low level of CEA

might be due to its appearance in advanced stage of malignancy.

There is an increase in nutrients demand in cancer cells for their growth, thereby tumour tissue

needs increase in blood supply. Therefore in cancerous tissue angiogenesis is required for the

progression of cancer. Increase in blood vessels needs increase in Hb for oxygen supply (Farooq

et al., 2002). Therefore in DMBA treated group Hb% level was increased significantly. Whereas

in disease treated with CRUEL CAPSULE Hb% level was found to be reduced, which indicates

the possible anti angiogenic property of CRUEL CAPSULE.

In our study the histopathologic evaluation showed that hyperplastic and well demarcated tubular

adenoma, the tubular and alveolar epithelium shows diffuse hyperplasia with minimal rim of

scletan muscle in disease control group, whereas in disease treated with CRUEL CAPSULE

there is no sign of inflammation or tissue damage, erosions / hyperplasic / neoplastic lesions was

observed. The resuls showes that the CRUEL CAPSULE produced antitumer activity by

inhibition of hyperplasic or neoplastic lesions thereby progress of the disease and metastasis.

21

cruel capsule - virgo · pdf filepharmacological evaluation of cruel capsule on rat mammary...

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