antianxiety effect of aqueous extract of fruits of emblica officinalis in rat

7/30/2019 Antianxiety Effect of Aqueous Extract of Fruits of Emblica Officinalis in Rat

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Pemminati S et al. / Jour nal of Pharmacy Research 2010, 3(2),219-223

Jour nal of Pharmacy Research Vol .3.I ssue 2.February 2010 219-223

Research Article

ISSN: 0974-6943 Available online through

www.jpronline.info

*Corresponding author.Sudhakar Pemminati,

Lecturer, Department of Pharmacology,Kasturba Medical College, Manipal University, Mangalore.India

Tel.: + 91-0824-2422271, extn: 5568,9448723549(M)

Telefax: +91--0824-2428183

E-mail :[email protected]

INTRODUCTION

Antianxiety effect of aqueous extract of fruits ofEmblica offi cinali s(EO) on acute and chroni

administration in rats.Pemminati Sudhakar, Gopalakrishna HN, Swati B, Shreyasi C, Pai MRSM, Vinod Nair1Department of Pharmacology, Kasturba Medical College, Manipal University,Mangalore-575 001, karnataka,India.Received on: 27-09-2009; Revised on: 16-11-2009; Accepted on:07-01-2010

ABSTRACT

The present study was undertaken to evaluate the anti-anxiety activity of aqueous extract of fruits ofEmblica officinalis (EO) on acute an

chronic administration in rats. Male Wistar albino rats weighing 150-180gms were divided into five groups (n=6) for acute and chronic stud

separately. Diazepam (1.0mg/kg) and EO (0.8, 2.0 & 4.0 mg/kg) were suspended in 1% gum acacia and administered orally. In acute stud

vehicle/drugs were given 60 minutes before, while in chronic study they were given once daily for 10 days and the last dose was given 60 mi

prior to exposure to the experimental paradigms viz., elevated plus maze and bright and dark arena. EO significantly increased the number

entries into, time spent and rears in the open arms. Also, it increased the percentile ratio of open arm to total arm entries in elevated plus maz

paradigm both on acute and chronic administration. In bright and dark arena paradigm the EO both on acute and chronic administration significantly increases the number of entries, time spent and rears in the bright chamber, also it reduced the duration of immobility. The

behavioural changes were comparable to that produced by the standard anxiolytic drug diazepam. The behavioural disinhibition produced b

EO suggestive of its anxiolytic like activity.

K e y w o r d s : Emblica officinalis, elevated plus maze, bright and dark arena.

Anxiety is a normal emotional behaviour. When it is severe

and/or chronic, however, it becomes pathological and can precipitate

or aggravate cardiovascular and psychiatric disorders. Although many

drugs are available in allopathic medicine to treat anxiety disorders,

they produce various systemic side effects or exhibit tolerance upon

chronic use. Benzodiazepines (BZDs) are the major class of com-

pounds used in anxiety and they remain the most commonly pre-

scribed treatment for anxiety. However the BZDs have many unwanted

side effects has prompted many researchers to evaluate new com-

pounds in the hope of identifying other anxiolytics with fewer side

effects 1. In ayurvedic medicine, many plant products have been

claimed to be free from side effects and less toxic than synthetic

drugs 2.

Embl ica of ficinali s Gaertn. (Phyl lanthus embl ica L.)(Family:Euphorbiaceae), popularly known as amla is a common house-

hold remedy that finds use in Indian indigenous system of medicine

against several ailments. Its fruits have been reported to possess

memory enhancer3, antiulcer4 ,hypoglycaemic5, hepatoprotective

hypolipidemic 7, antipyretic and analgesic 8, anti-inflammatory9, ant

bacterial10, immunomodulatory 11, antioxidant12, gastroprotective

activity on central nervous system14 and anticataleptic activity15. Th

aqueous extract of fruits of Emblica officinalis contains 30.0%

Tannins & 13.4% of Gallic acid (estimation and purity of active pri

ciple was done by the Quality Control Laboratory, M/s. Natural Rem

edies, Bangalore, lab reference no.0505211, dt.31-05-2005).

From our laboratory, we have reported the anxiolytic effe

of NR-ANX-C,a polyherbal product in rats16. NR-ANX-C is com

posed ofWithania somnifera , Ocimum sanctum, Camellia sinensi

Emblica officinallis and Shilajit. As EO is one of the componen

and its antianxiety activity is not reported so far. In order to evalua

the contribution or role of EO in anxiolytic activity, the present studwas undertaken. Two pharmacologically validated experimental mo

els, elevated plus maze 17 and bright and dark box18 were employed

MATERIALS AND METHODS

Animals:

Male Wistar albino rats weighing 150-180g (90-110 days olwere used for the study. They were housed in clean, clear polyproplene cages in groups of four in each cage maintained at 24 50C with

hours light and dark cycle with free access to food and water. The an

mals were acclimatized to laboratory conditions before testing. Ex


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periments were conducted between 9 00 to14 00 hours. Each rat was

used only once. The study was approved by the Institutional Animal

Ethical Committee (IAEC) and the study was conducted according to

the Indian National Science Academy guidelines for the use and care

of experimental animals.

Drugs and ChemicalsThe standard anxiolytic, diazepam (Ranbaxy Pvt ltd., Mumbai)

and the test drug, a dry powder of aqueous extract of fruits ofEmblica

officinalis (Natural Remedies, Bangalore) were suspended in 1% gum

acacia solution19. Each drug solution was prepared freshly just before

the administration. Drugs and vehicles were administered orally. The

doses of each drug were selected on the basis of earlier finding with

NR-ANX-C16. Control group of animals were received appropriate

volume of vehicle, 1% gum acacia solution. Drugs, dosage and num-

ber of animals used per treatment were shown in (Table 1).

Table 1. Groups, treatment and doseGroups Treatment

( n = 6)I Control- 1%Gum acacia - 10.0ml/kgII Diazepam - 1.0mg/kgIII Emblica officinalis 0.8 mg/kgIV Emblica officinalis - 2.0 mg/kg

V Emblica officinalis 4.0 mg/kg

In acute study, drugs/vehicle was administered 60 min prior

to experiment. In chronic study they were administered once daily for

10 days and the last dose was given on the 10th day, 60 min prior to

experiment.

Apparatus

Elevated plus maze

The wooden maze consisted of two open arms (length 50 cmX breadth 10 cm) and two closed arms of the same size (height 40 cm).

The arms of the same type were opposite to each other, with a central

square of 10 cm. The maze was elevated to a height of 50 cm above the

floor17.

Bright and dark box

The apparatus consisted of an open top wooden box. Two

distinct chambers, a black chamber (20 X 30 X 35 cm) painted black

and illuminated with dimmed red light and a bright chamber (30 X 30 X

35 cm) painted white and brightly illuminated with 100 W white light

sources, were located 17 cm above the box. The two chambers were

connected through a small open doorway (7.5 X 5 cm) situated on the

floor level at the centre of the partition18.

Behavioral assessment.

Each animal was tested initially in plus maze and, then, in

bright and dark arena paradigm in a single setting. In acute study 60

min after and in chronic study 60 min after the last dose on the 10th

day of drug or vehicle administration, each animal was placed in the

centre square of the plus maze, facing one of the open arms. The

number of entries into and the time spent in open and closed arms and

the number of rears in each arm in a five-minute period was noted.

Following the elevated plus maze test, the animal was placed at th

centre of the brightly lit arena in the light and dark box. The number

entries into and the time spent in the bright arena, the number of rear

in the bright and dark arenas and the duration of immobility wer

noted. Following each trial, the apparatus were cleaned to mask th

odour left by the animal in the previous experiment. Hand operate

counters and stop watches were used to score the behaviour of anmals.

Statistical Analysis:

All results were expressed as mean standard error (SEM

and analysed by one-way ANOVA with drug treatment as the ind

pendent factor. Post-hoc comparisons were performed by applyin

Dunnets test.P


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b. Bright and dark arena

Acute Study:

Diazepam (1.0mg/kg) treated rats significantly increased the

number of entries into the bright arena the time spent and the rears in

bright arena. EO at the highest dose tested(4.0mg/kg) significantly

increased the number of entries into, time spent and rears in bright

arena and decreased the duration of immobility. However increasetime spent was significant at all the doses tested (0.8,2.0 &4.0mg/

kg)[Table - 4]

Chronic Study:

Repeated administration of diazepam (1.0 mg/kg) significantly

increased the time spent and the rears in light arena and decreased the

duration of immobility. However, increase in the number of entries

into bright arena was not significant. EO at all the doses tested(0.8,2.0

& 4.0mg/kg) increased the time spent in bright chamber, number of

rears in dark chamber and decreased duration of immobility, The ex-

tract also increased the the number of rears at higher doses( 2.0 and

4mg/ kg)and the number of entries in bright chamber at the highest

dose, 4mg/kg. [Table - 5]

DISCUSSION:

The two experimental models of anxiety, elevated plus maze

and bright and dark arena, are based on the assumption that unfamil-

iar, non-protective and brightly lit environmental stress provokes in-

hibition of normal behaviour. This normal behavioural inhibition is

further augmented in the presence of fear or anxiety like state.

In the elevated plus maze, the open arms are more fear pro-

voking than the closed arms. The ratio of entries, time spent and rear-

ing behaviour in open arms to closed arms reflects the safety of closed

arms with relative fearfulness of open arms 17. The reduction in entry,

time spent, rearing in open arms, ratio of open arm to total arm entries

and increased defecation are the indications of high level of fear or

anxiety. Anxiolytic drugs increase the proportion of entries, time spent

and rearing in open arms. They also increase the ratio of open arm to

total arm entries.

In the bright and dark box paradigm, the brightly lit environ-

ment is a noxious environment stressor that inhibits the exploratory

behaviour of rodents. Reduction in the number of entries, time spent

and rearing behaviour in the light chamber is regarded as markers of

anxiety18. Rearing reflects an exploratory tendency20 of the animal that

can be reduced due to a high level of fear.

An increase in the number of entries to open arm , time spent and rears

in open arms, and percentile ratio of open to total arm entries in

elevated plus maze paradigm and increase in the number of transitionto , time spent and rears in bright arena in bright and dark arena para-

digm by the rats treated with standard anxiolytic diazepam suggest

the decreased fear, decreased aversion to light and increased explor-

atory behaviour21. The test compound, Emblica officinalis ,also pro-

duced the behavioural changes in both paradigms similar to that

produced by diazepam indicating that the test drug has behavioural

disinhibitory or anxiolytic like activity.The anxiolytic effect of EO was

consistent with our earlier report of anxiolytic effect of NR-ANX-C16,

in which EO is one of the components.

Despite the wide spread traditional use of EO for treatin

various disorders, there are no reports of any scientific evaluation o

its antianxiety effect. Earlier reports on the chemical constituents o

plants and their pharmacology suggest that plants containing fla

vonoids, saponins and tannins possess activity against many CN

disorders22. Further studies are required to elucidate the possiblmechanism of anxiolytic activity and its usefulness in human being

ACKNOWLEDGMENTS.

The authors are grateful to Natural Remedies Pvt. Ltd, Ban

galore for providing the aqueous extract of fruits ofEmblica officinal

powder.

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antianxiety effect of aqueous extract of fruits of emblica officinalis in rat

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