Imperial J.

Pharmacology & Toxicology 1(1): June 2011

ISSN: 2248 9746

Imperial Journal of Pharmacology & Toxicology

www.imperialpharmajournals.com [Research Article] --------------------------------------------------------------------------------------------------------------------------------------------------

THE PHARMACOLOGICAL EVALUATION OF FOENICULUM VULGARE MILLER FOR ANTIANXIETY

Archana Divekar*1, Rajesh J. Oswal2, Yogita R. Bagul1, Rishikesh V. Antre2
1 2

Department of Pharmacology, JSPMS Charak college of Pharmacy and Research, Wagholi, Pune, MH, India Department of Pharmaceutical Chemistry, JSPMS Charak college of Pharmacy and Research, Wagholi, Pune, MH, India

ABSTRACT Correspondence to Author Ms. Archana Divekar Department of Pharmacology, JSPMS Charak college of Pharmacy and Research, Wagholi, Pune, MH, India Email jspmpharmacy@gmail.com Key Words Foeniculum vulgare Miller, Anti-anxiety activity, Elevated plus-maze model, Diazepam. Foeniculum vulgare Miller is commonly used in India as flavouring agent for food preparation. The crude ethanolic extract of Foeniculum vulgare Miller contains Anisic acid, Anisic aldehyde, Dpinene, Fenchone, organic Phellandrine, Volatile oils (50-60%) Anethole. Since the crude extract of Foeniculum vulgare Miller contains aromatic oil as revealed by phytochemical assay. Here it was decided to explore the anxiolytic activity on lab animal. The crude ethanolic extract of Foeniculum vulgare Miller has been explored for its anxiolytic activity on albino mice by Elevated plusmaze model. The results were very encouraging. The extract at doses of 200 mg/kg and 400 mg/kg has been found to possess significant anti-anxiety activity on the tested experimental models. The extract (400 mg/kg) exhibited maximum anti-anxiety effect. At a higher dose the extract (400 mg/kg) showed increase number of entries and time spent in open arm of Elevated plus-maze model. The effect produced by the extract was comparable to that of Diazepam, a prototype of Anxiolytic agent.

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Imperial J. Pharmacology & Toxicology 1(1): June 2011

ISSN: 2248 9746

INTRODUCTION It has many biological activities like antibacterial, anti-inflammatory, antispasmodic and stimulating activity. It was found that the aromatic oil is the main constituent responsible for stimulant effect. Anxiety is a psychological and physiological state characterized by cognitive, somatic, emotional, and behavioral components1. These components combine to create an unpleasant feeling that is typically associated with uneasiness, fear, or worry. Types of anxiety disorder are as follows 1. Panic disorder. 2. Panic disorder with agoraphobia 3. Generalized anxiety disorder. 4. Social anxiety disorder/social phobia. 5. Post-traumatic stress disorder (PTSD). An anxiolytic (also antipanic or antianxiety agent2) is a drug used for the treatment of symptoms of anxiety. Anxiolytics have been shown to be useful in the treatment of anxiety disorders. Anxiolytics are also known as "minor tranquilizers3" though their use and effects are by no means minor; this term is less common in modern. Types of Anxiolytics 1. Benzodiazepines 2. Azapirones 3. Barbiturates 4. Hydroxyzine 5. Herbal treatments Types of behavioural model for anxiolytic agents a) Exteroceptive stimuli models b) Interoceptive stimuli models a) Exteroceptive stimuli models 1) Response-independent presentation of stimuli. Open field test. Y-maze model. Elevated plus-maze model. Head dipping test. Black and white test box. 2) Response-contingent presentation of stimuli. Geller-seifters test. Vogel conflicts test. Quinteros punished bar pressing. Available online on: www.imperialpharmajournals.com

Social interaction test. b) Interoceptive stimuli models Electrical stimulation of brain. Pharmacological manipulation (drug discrimination tests) a) FG-7142-induced anxiety. b) Caffeine induced anxiety. c) Yohimbine-induced anxiety. d) Flumazenil-induced anxiety. e) Amphetamine-induced anxiety. Aggression / anxiogenesis a) Foot shock-induced Aggression. b) Drug-induced Aggression. c) Isolation-induced Aggression. We are selected elevated plus maze model because, it is simple and less time consuming procedure, does not involve any sophisticated equipment nor prior training nor noxious stimuli. Based on spontaneous behavior of the animal, the test provides a valid and reliable measure of anxiety in animals and for testing of antianxiety drugs4. Elevated Plus-Maze Model (EPM) The elevated plus maze (EPM) is a rodent model of anxiety that is used as a screening test for putative anxiolytic compounds and as a general research tool in neurobiological anxiety research.

The test setting consists of a plus-shaped apparatus with two open and two enclosed arms, each with an 17

Imperial J. Pharmacology & Toxicology 1(1): June 2011

ISSN: 2248 9746

open roof, elevated 4070 cm from the floor5. The model is based on rodents' aversion of open spaces. This aversion leads to the behavior termed thigmotaxis, which involves avoidance of open areas by confining movements to enclosed spaces or to the edges of a bounded space. In EPM this translates into a restriction of movement to the enclosed arms. Anxiety reduction in the plus-maze is indicated by an increase in the proportion of time spent in the open arms (time in open arms/total time in open or closed arms), and an increase in the proportion of entries into the open arms (entries into open arms/total entries into open or closed arms). Total number of arm entries and number of closed-arm entries are usually employed as measures of general activity. While EPM is the most commonly employed behavioral animal anxiety model, there are several issues concerning the validity of the model. Classical clinical anxiolytics, such as benzodiazepines (e.g., Valium), do reduce measures of anxiety in EPM. However, more novel compounds, such as 5-HT1A agonists (e.g., Buspar) give mixed results. Selective serotonin reuptake inhibitors and tricyclic antidepressants, which are commonly employed in clinical settings to treat anxiety disorders, also do not lead to a stable anxiolytic effect on EPM. This raises the possibility that EPM is a suitable model for testing GABA-related compounds, such as benzodiazepines or direct GABAA agonists, but not for other drugs. Despite this, the model is commonly employed for screening putative anxiolytics and for general research into the brain mechanisms of anxiety, because of the ease of employment and the vast number of studies already in the literature6,7. Material and Method Plant Material The plant was collected in November 2009 from the Wagholi, Pune district of Maharashtra, India. The plant was taxonomically identified by the Botanical Survey of India, as Foeniculum vulgare Miller. The fruits were dried under shade with occasional shifting then powdered with a mechanical grinder and stored in an airtight container. Preparation of extract Available online on: www.imperialpharmajournals.com

The powder obtained was subjected to successive soxhlet extraction with the solvents with increasing order of polarity i.e. Ethanol. Extraction were carry out for 72-80 hrs by standard Soxhlet apparatus method and the extracts were evaporate to determine the percentage yield and Phytochemical screening of plant. Animal Male Albino mice (Swiss strain) weighing 25-30g, were be house under standard laboratory condition, in a group of six each. The animals were having free access to food and water at liabdum as per norms ethical committee and CPCSEA norms of the institute approved the protocol of the study. For this study we used Diazepam as standard drug. Method Swiss albino mice weighed (25-30 g) and numbered the animal. Divided them into four groups each consisting of 6 mice: a) Group 1. As a normal control (saline water). b) Group 2. Received standard drug (Dose: 2 mg/kg by I.P.) c) Group 3. Received dose of extract (Dose: 200 mg/kg by orally) d) Group 4. Received dose of extract (Dose: 400 mg/kg by orally) Placed the animal individually in the centre of the maze, head facing towards open arms and start the stop watch and note following parameters for five minutes: a) First preference of mouse to open or enclosed arm. b) Number of entries in open arm and enclosed arm (an arm entry defined as the entry of four paws into the arm). c) Average time each animal spends in each arm (average time =total duration in the arm / number of entries). Inject diazepam to the standard group. After 30 min place the animal individually in the centre of the maze and note the all parameters open and closed arm entries and time spend in open and closed arm. Compare the preference of the animal to open /enclosed arm, average time spent in open arm number of entries and average time spent by the mouse in the open arm.

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Imperial J. Pharmacology & Toxicology 1(1): June 2011

ISSN: 2248 9746

Results 1) 2) Table no.1: Phytochemical screening of Foeniculum Vulgare Miller

Sr. no. 1. 2. 3. Constituents Carbohydrate Volatile oil Glycosides Tests Molisch reagent Fehlings reagent Solubility test Filter paper test Coumarin test Flavonoid test 4. Tannins and phenolic compounds Lead acetate test Fecl3 test Potassium dichromate Biuret test Millions test Xanthoprotein test Test for vitamin c Ethanol Extract + + + + + + + + + + + + +

5. 6.

Protein Vitamins

Acute toxicities study The lethal dose (LD50) has been found in the selected extract of Foeniculum vulgare Miller plant on the selected albino mice. We got the of LD50 range 2000 mg/kg body weight. As per the Lethal dose (LD50) we have selected effective dose (ED50) 200mg/kg lower dose and 400mg/kg higher dose of ethanolic crude extract of Foeniculum vulgare Miller. Anti-anxiety Diazepam (2 mg/kg) treated mice showed a significant increases in the number of open arm entries and time

spend in open arms. And they showed reduction in time spend in the closed arms. An ethanolic extract of F. Treated mice exhibited significant increases in number of open arm entries, time spend in open arms and percentile ratio of open arms to total arm entries at both selected dose. (200 & 400 mg/kg) The result was significant of higher dose of ethanolic extract at **p< 0.01 for Elevated plus Maze method. Anti-anxiety activity is presented in following table

Table No. 2 Anti-anxiety activity of Founiculum vulgare Miller by Elevated Plus Maze Model Sr. No. Groups (Dose mg/kg) 1 2 3 4 No. of entries % present to Average time spent open arm Open Closed 25 09.777.30 29.002.11 84 32.670.21 8.341.89 57** 20.340.76* 16.22.231.32 76*** 28.110.80** 13.980.54

Open Closed Control 2.100.230 6.250.453 Standard(Diazepam) 12.340.564 2.220.32 Extract (200 mg) 6.570.768* 4.880.129 Extract (400 mg) 9.670.433** 3.010.441

(Values are expressed as mean + S.E.M. n=6 *** p < 0.001, ** p < 0.01, * p < 0.05 compared with vehicle control ANOVA followed by Dunnets t- test). Available online on: www.imperialpharmajournals.com

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Imperial J. Pharmacology & Toxicology 1(1): June 2011

ISSN: 2248 9746

The anti-anxiety of an ethanolic extract of fruits of Foeniculum vulgare Miller (200&400 mg/kg orally) both doses lower and higher respectively is comparable with standard drug Diazepam (2 mg/kg). There is an increase in open entries and time spent in open arm as exhibited by the extract when compared with the control group. It is computed in graph:1 which shows the effectiveness and pharmacological rationale for the use of Foeniculum vulgare Miller as an anti-anxiety drug. REFERENCES 1. Seligman, M.E.P., Walker, E.F. & Rosenhan, D.L. (2001). Abnormal psychology, (4th ed.) New York: W.W. Norton & Company, Inc. 2. Antianxiety agent at Dorlands Medical Dictionary.

3. "WordNet Search 3.0". http://wordnetweb.princeton.edu/perl/webwn?s=anxio lytic. Retrieved 2009-01-01. 4. S.K. Kulkarni, Handbook of experimental pharmacology, vallabh prakashan, Third edition, page no.30 . 5. Kokate C.K., Purohit A.P., Gokhale S.P., textbook of Pharmacognosy, Nirali Prakashan, 2008, Page no.334. 6. Khandelwal K.R., Experimental & Techniques of practical pharmacognocy, 19th edition Nirali prakashan, Pune 2008. Page no. 149-155. 7. Barlow, David H. (November 2002). "Unraveling the mysteries of anxiety and its disorders from the perspective of emotion theory". American Psychologist: 124763. 8. Ohman, A. (2000). Fear and anxiety: Evolutionary, cognitive, and clinical perspectives. In M. Lewis & J. M. Haviland-Jones (Eds.). Handbook of emotions. (pp.573-593). New York: The Guilford Press.

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