Evaluation of antidepressant activity of clitoris ternatea in animals

Evaluation of antidepressant activity of clitoris ternatea in animals

1. “EVALUATION OF ANTI DEPRESSANT ACTIVIY OF CLITORIA TERNATEA IN ANIMALS” Under the guidance of Mr.Sunil Kumar M.Pharm (PhD)

2. Presented By :- Akhila Reg.No.19P2506 Aniket Reg.No.19P2508 Halbarge Sushma Reg.No.19P2516 Sukanya Biradar Reg.No.19P2550 Suma D Jyoti Reg.No.19P2551

3. CONTENTS INTRODUCTION AIMS AND OBJECTIVES MATERIALS AND METHODS RESULTS DISCUSSION SUMMARY CONCLUSION

4. Depression is highly prevalent condition, affecting approximately 121 million people worldwide according to WHO 2008. It is one of the most prevalent and costly psychiatric disorders worldwide It is the fourth leading cause of disability worldwide. Major depression and mania are two extremes of affective disorder which refer to pathological change in mood state. Although there are many effective antidepressants available today. The current armentariam of therapy is often inadequate with unsatisfactory results in about one third of all subjects treated. INTRODUCTION

5. This necessities the development of newer and more effective antidepressant from traditional medicinal plants whose psychotherapeutic potential had been assessed in variety of animal models. Herbal drugs play important role in healthcare programs in treatment of various diseases including depression disorders. Plant kingdom is rich of various medicinal species having their effect on nervous system. Now a day’s pharmacological spectrum and biological efficiency of such herbal drugs can be suitably established due to development of various neuropharmacological testing of herbal drugs with effect on CNS and ANS. Keeping in view the above ideas, this work was planned on CLITORIA TERNATEA plant extracts in animal models of experimentally induced depression.

6. OBJECTIVES The present study aims to investigate antidepressant activity of Clitoria Ternatea leaf extracts in animal models of acute depression. Objectives of the study: Preparation of Successive solvent extraction of chloroform, alcoholic and aqueous extract in increasing polarities of Clitoria Ternatea . To perform phytochemical screening of chloroform, alcohol and aqueous extracts on leaves of Clitoria Ternatea . To assess the acute toxicity of Clitoria Ternatea leaves extracts using mice as animal model (OECD guidelines -420). To record and compare dose dependent antidepressant activity in vehicle treated, test treated and standard antidepressant drug in acute animal models of depression.

7. CLITORIA TERNATEA Clitoria Ternatea belongs to Fabaceae family. Clitoria Ternatea commonly known as Asian pigeon wings, blue bellvine , blue pea, butterfly pea, or Darwin pea In India it is reversed as a holy flower used in daily puja rituals. Kingdom : Plantae Order : Fabales Family : Fabaceae Subfamily : Faboideae Genus : Clitoria Species : C.ternata MATERIALS AND METHODS

8. Selections of Animals Healthy Albino Swiss mice of either sex weighing between 18-25gms. Albino Wistar rats of either sex, weighing between 160-180gms were employed for study. MATERIALS AND METHODS

9. Phytochemical analysis Preliminary qualitative tests The extracts were subjected to preliminary qualitative phytochemical investigation . Determination of acute oral toxicity The acute oral toxicity study of the extract was carried out by using Albino Swiss mice of either sex weighing between 18-25 g as per OECD (Organization for Economic Cooperation and Development) Guidelines 420. Low and high dose of test drug was selected for the treatment. MATERIALS AND METHODS

10. Experimental Models RAT FORCED SWIM TEST (FST) TAIL SUSPENSION TEST (TST) LOCOMOTOR ACTIVITY MATERIALS AND METHODS

11. RESULTS Preliminary phytochemical testing of Clitoria Ternatea extracts: The chemical constituents of various extracts of Clitoria Ternatea indicated the presence of following constituents : Chloroform extract -flavonoids, reducing sugar, starch, tannin and phenolic compounds. Ethanol extract – carbohydrates, starch, gum, flavonoids, steroid, saponin glycosides, reducing sugar, tannin, alkaloid and phenolic compound. Aqueous extract -flavonoids, carbohydrates, starch, gum, reducing sugar, proteins, saponin glycosides, tannin and phenolic compound.

12. FORCED SWIM TEST Comparative profile of immobility parameter in rat forced swim test after acute treatment of 200mg/kg and 400mg/kg of Ethanol Extracts of Clit oria Ternatea All values are mean ± SEM in sec. (n=6).One-way ANOVA followed by Dunnet ’ s test. * P < 0.05, ** P <0.01, *** P <0.001, when compared to vehicle treated animals Group Treatment Mean duration of immobility(sec) 1 Control (0.2ml/animal) 193±9.977 2 Imipramine(15mg/kg) 81±22.38** 3 CTEe-1(200mg/kg) 123.8±22.02* 4 CTEe-2(400mg/kg) 73±16.96***

13. Comparative profile of immobility parameter in rat forced swim test after acute treatment of 200mg/kg and 400mg/kg of ethanol extracts of C litoria Ternatea

14. FORCED SWIM TEST Comparative profile of immobility parameter in rat forced swim test after acute treatment of 200mg/kg and 400mg/kg of aqueous extracts of C litoria Ternatea All values are mean ± SEM in sec. (n=6).One-way ANOVA followed by Dunnet ’ s test. * P < 0.05, ns=Nonsignificant when compared to vehicle treated animals Group Treatment Mean duration of immobility(sec) 1 Control (0.2ml/animal) 176±17.98 2 Imipramine(15mg/kg) 85±26.36* 3 CTAe-1(200mg/kg) 133.2±27.19ns 4 CTAe-2(400mg/kg) 85.17±18.11*

15. Comparative profile of immobility parameter in rat forced swim test after acute treatment of 200mg/kg and 400mg/kg of aqueous extracts of C litoria Ternatea Mean duration of immobility(sec)

16. FORCED SWIM TEST Comparative profile of immobility parameter in rat forced swim test after acute treatment of 200mg/kg and 400mg/kg of chloroform extracts of All values are mean ± SEM in sec. (n=6).One-way ANOVA followed by Dunnet ’ s test. ** P <0.01, ns= Non significant when compared to vehicle treated animals. Group Treatment Mean duration of immobility(sec) 1 Control (0.2ml/animal) 184.5±8.958 2 Imipramine(15mg/kg) 111.0±24.61** 3 CTCe-1(200mg/kg) 183.2±16.29 ns 4 CTCe-2(400mg/kg) 173.3±9.218 ns

17. Comparative profile of immobility parameter in rat forced swim test after acute treatment of 200mg/kg and 400mg/kg of chloroform extracts of C litoria Ternatea

18. TAIL SUSPENSION TEST Comparative profile of immobility parameter in mice tail suspension test after acute treatment of 200mg/kg and 400mg/kg of ethanol extracts of Clitoria Ternatea All values are mean ± SEM in sec. (n=6).One-way ANOVA followed by Dunnet ’ s test. * P < 0.05, *** P <0.001, when compared to vehicle treated animals. Group Treatment Mean duration of immobility(sec) 1 Control(0.2ml/animal) 137.7 ± 14.96 2 Imipramine(15mg/kg) 60.33 ± 4.310*** 3 CTEe-1(200mg/kg) 96.67 ± 10.56* 4 CTEe-2(400mg/kg) 65.67 ± 12.42***

19. Comparative profile of immobility parameter in mice tail suspension test after acute treatment of 200mg/kg and 400mg/kg of ethanol extracts of Clitoria Ternate a

20. TAIL SUSPENSION TEST Comparative profile of immobility parameter in mice tail suspension test after acute treatment of 200mg/kg and 400mg/kg of aqueous extracts of Clitoria Ternatea All values are mean ± SEM in sec. (n=6).One-way ANOVA followed by Dunnet ’ s test. * P < 0.05, ** P <0.01, when compared to vehicle treated animals. Group Treatment Mean duration of immobility(sec) 1 Control (0.2ml/animal) 150.2 ± 14.63 2 Imipramine(15mg/kg) 70.33 ± 16.95** 3 CTAe-1(200mg/kg) 80.17 ± 14.23* 4 CTAe-2(400mg/kg) 77.83 ± 16.81*

21. Comparative profile of immobility parameter in mice tail suspension test after acute treatment of 200mg/kg and 400mg/kg of aqueous extracts of Clitoria Ternatea

22. TAIL SUSPENSION TEST Comparative profile of immobility parameter in mice tail suspension test after acute treatment of 200mg/kg and 400mg/kg of chloroform extracts of Clitoria Ternatea All values are mean ± SEM in sec. (n=6).One-way ANOVA followed by Dunnet ’ s test. * P < 0.05, ** P <0.01, ns= Nonsignificant, when compared to vehicle treated animals. Group Treatment Mean duration of immobility(sec) 1 Control (0.2ml/animal) 149.8±10.14 2 Imipramine(15mg/kg) 68.83 ± 11.46** 3 CTCe-1(200mg/kg) 95.83 ± 12.00* 4 CTCe-2(400mg/kg) 122.3 ± 84.00ns

23. Comparative profile of immobility parameter in mice tail suspension test after acute treatment of 200mg/kg and 400mg/kg of chloroform extracts of Clitoria Ternatea

24. LOCOMOTOR ACTIVITY Comparative profile of change in locomotor activity in rats after acute treatment of ethanol extract of Clitoria Ternatea All values are mean ± SEM in sec. (n=6).One-way ANOVA followed by Dunnet ’ s test. ns= Nonsignificant, when compared to vehicle treated animals. Group Treatment Locomotor activity scores 1 Control (0.2ml/animal) 345.83±26.76 2 Imipramine(15mg/kg) 356.66±34.84 3 CTEe-1(200mg/kg) 343.16±17.97ns 4 CTEe-2(400mg/kg) 361.1±33.06ns

25. Comparative profile of change in locomotor activity in rats after acute treatment of ethanol extract of Clitoria Ternatea

26. LOCOMOTOR ACTIVITY Comparative profile of change in locomotor activity in rats after acute treatment of Aqueous extract of Clitoria Ternatea All values are mean ± SEM in sec. (n=6).One-way ANOVA followed by Dunnet ’ s test. ns= Nonsignificant, when compared to vehicle treated animals. Group Treatment Locomotor activity scores 1 Control (0.2ml/animal) 362.7±29.84 2 Imipramine(15mg/kg) 356.16±22.07 3 CTAe-1(200mg/kg) 354.16±24.87ns 4 CTAe-2(400mg/kg) 358.83±28.23ns

27. Comparative profile of change in locomotor activity in rats after acute treatment of aqueous extract of Clitoria Ternatea

28. LOCOMOTOR ACTIVITY Comparative profile of change in locomotor activity in rats after acute treatment of Chloroform extract of Clitoria Ternatea All values are mean ± SEM in sec. (n=6).One-way ANOVA followed by Dunnet ’ s test. * P < 0.05, ns= Nonsignificant, when compared to vehicle treated animals. Group Treatment Locomotor activity score 1 Control (0.2ml/animal) 261.3±22.95 2 Imipramine(15mg/kg) 257.83±16.78 3 CTCe-1(200mg/kg) 169.0±29.36* 4 CTCe-2(400mg/kg) 162.8±16.39*

29. Comparative profile of change in locomotor activity in rats after acute treatment of chloroform extract of Clitoria Ternatea

30. DISCUSSION The main finding of present investigation suggests the antidepressant activity of extracts of Clitoria Ternatea leaves in rat forced swim test, tail suspension test in mice and open field test in rats. In phytochemical study, observed the presence of flavonoids (flavonol glycosides) in Clitoria Ternatea leaves extracts. Recently several studies have suggested the antidepressant effect of flavonol glycosides. Therefore one of the antidepressant mechanisms of Clitoria Ternatea leaves extracts is thought to involve flavonoids, which exerts an antidepressant effect. Acute administration of Clitoria Ternatea leaves extracts demonstrated significant (compared to vehicle treated group) dose dependent reduction in duration of immobility.

31. DISCUSSION In forced swim test, The effect of 400mg/kg (CTEe-1) was better than 15mg/kg imipramine (TCA).The effect of 200mg/kg (CTEe-2) was significant when compared to vehicle treated group but was not better than 15mg/kg imipramine treated animals. In tail suspension test, acute treatment of CTAe both 200mg/kg and 400mg/kg shown significant dose dependent reduction in duration of immobility but did not produce acute effect better than 15mg/kg imipramine treated animals. In Locomotor activity, acute treatment dose dependent effect was observed in CTCe treated animals. There was significant (compare to vehicle treated animals) reduction in locomotor activity at dosage 200mg/kg and 400mg/kg compared to vehicle treated animals in open field test (Actophotometer).

32. . The acute toxicity study were conducted as per OECD guideline 420. It was found that the ethanol, aqueous and chloroform extracts even at 200mg/kg dose had not shown any mortality. Hence, it confirms that it is practically non toxic in nature. Ethanol and aqueous extracts of Clitoria Ternatea has dose dependent antidepressant activity at low and high dose comparable to standard imipramine drug in test. Chloroform extract of Clitoria Ternatea has significant antidepressant activity at low dose comparable to imipramine in TST. It is clear that the ethanol and aqueous extracts of Clitoria Ternatea had no influence on rat locomotor activity but chloroform extract of Clitoria Ternatea decrease the locomotor activity. CON CLUSION & SUMMARY