Neuropharmacological screening of Ficus Carica Linn; Fruit for Anxiolytic and Antidepressant Activity

Background: Rutin and polyphenol which are present in ficus carica linn are responsible for anti-depressant and anxiolytics effect. Aims: We have conducted the study to evaluate the anti depressant and anxiolytic effect of ficuscaricalinn on mice and compare its effect with well known prescribed anti depressant and anxiolytic drug alprazolam. Methods: Three groups of mice were marked as Control, Reference and Test. Each group comprise of 5 mice. Control group was on normal diet. Reference group was feeded alprazolam at a dose of 0.5mg/60kg while test group were provided two different dilution of ficuscaricalinn, one dilution is 250mg/ kg/100ml of water and second dilution is 500mg/kg/100ml of water. We used Light/dark box method and Elevated plus maze method to evaluate the anxiolytic effect of ficus carica linn. To determine the anti-depressant effect of ficus carica linn. Results: we was used one method that is Head Dip Method. Results were collected and observed readings and data were clearly indicating the great and pronounced anti depressant and anxiolytic effect of ficus carica linn at a dose of 500mg/kg compare with a widely prescribed drug alprazolam for its anti-depressant and anxiolytic effect.Ficus carica is well known for its numerous biological activities also found to be one of excellent distressing agent. Stress provoking several disorders could be limit by the use of ficus carica fruit in doses 250mg/kg and 500mg/kg. Ficus carica linn as a natural product may prove better therapeutic agent if more study is conducted on it. Conclusion: As this study is confined to the mice, there’s much possibilities to have similar effect in humans but require more work on it.


Introduction:
Anxiety & depression are amongst most prevailing mental or psychic disorder currently occurring worldwide. Anxiety is the tense condition with the feeling of displeases, worry and fear 1 . One suffers from agitation and nervousness in performing regular affairs of life. The frequently observed sign and symptoms been encounter in such individuals are dyspnea, tachycardia, xerostomia, uneasiness, muscle tightness and stress 2 .Although benzodiazepines are most extensively used and prescribed classes for anxiety and depression, 1 there are so many issues and complication associated with it. On a long term (regular) use, tolerance and dependence may develop by the body, chances of drug abuse may increase and adverse effects are appeared. The chronic use can also potentiate depression and may lead to suicide attempts in impulsive patient 3 .Moreover many short acting barbiturates are provided to the anxious patients to gain the benefit of its sedating activity 1 . These agents possess strong risk of drug toxicity and needs different methods for the proper evaluation of their plasma levels to subside its harmful toxic effects 4 .These drugs are cytochrome p450 inducer so possess numerous drug-drug interactions and the toxic doses are precipitated to coma 1 . Tricyclic antidepressants (TCAs) and selective serotonin re-uptake inhibitors (SSRIs) are recommended to patients complaining for pressure, anxiety and depression. TCAs are reserved for those individuals who didn't respond well to SSRIs. They owe high potential for decreasing blood pressure, increase in body weight and other anticholinergic effects such as blurred vision; dry mouth etc. 3 . Sexual dysfuntioning, insomnia, G.I upset and weight gain are typically observed with the use of SSRIs 5 .
According to the Royal Society of Medicine 15% of the global burden of disease is assigned to mental disorders, in high-income countries it is 30%, in middle-income countries its 20% and it is 10% in low-income countries. So probably anxiety is the problem facing all over the globe 6 . In the past centuries so many treatments were provided to get rid of anxiety, the investigation on passion fruit gives out positive results by exerting the antianxiety action at low doses, 7 similarly another research conducted on a traditional Asian herbal medicine yokukansan for determining its anxiolytic activity on old aged rats is also notable 8 . The literature established comprising of past research on animals and human clinical trials prove many dietary supplements possess mind relaxing and anxiolytic actions 9 . Hence these secondary cures to anxiety are cheap source of earning valuable agents that aims to reduce tension and stress 9 .
The ficus plant contains variety of bioactive constituents i.e. Coumarins, flavonoids 10.11 sterols, triterpenoids, anthocyanins 10 etc. which are very important as far as both nutritional and medicinal merits are concerned. Different parts of the plant could be use for several therapeutic actions including antihypercholesterolaemia 11 .antidiarrheal 13 , antibacterial 14 , antioxidant 11 , antiinflammatory 11 , anticancer 11 , antispasmodic 15 , antidiabetic 12,16 , anthelmintic 12 , antiplatelet 15 , anti-inflammatory 11, antipyretic 12, gastro protective 11 , immunodulatory 12 and hepatoprotective 10-12.17,18 Polyhydroxyphenols commonly known as polyphenols are sub classify into phenolic acids, flavonoids etc. A healthy diet consist of a fine good amount of naturally existing polyphenols render great activity against the oxidative stress which is one of the root cause of anxiety, depression and other central nervous system related diseases 19 . Rutin which is present in greatest amount in ficus fruit may act as an anxiolytic and relieves fatigue 20 .
Rutin found in mulberry leaf extract follows twocompartment model in rats, show rapid absorption and slow decreased in plasma levels 22 . Here are some phenolic of different concentrations in Ficus carica fruit obtained through HPLC-PDA system; rutin was found chiefly up to 28.7 mg per 100 g FW (fig weight), (+)-catechin chlorogenic acid , (−)-epicatechin, gallic acid, syringic acid were also found in concentrations upto (4.03 mg, 1.71 mg, 0.97 mg, 0.38 , 0.10 mg) per 100 g FW, respectively 21 . The study which evaluates the anxiolytic properties of aqueous acetonic extracts of Ficus carica L. leaves, we belief that the fruit could also have possibilities of certain central nervous system activities 6 as well as the presence of high concentration of phenols found might be responsible for anti stress effect. In favor of this assumption we have investigated the two different dilutions of different concentrations of ficus carica fruit by administering orally in mice adopting elevated plus maze method and light and dark box test for determination of anxiolytic action and hole-board apparatus for the determination of antidepressant action of ficus carica fruit.

Methodology: Animals:
Swiss mice (n=5) of both gender with average weight of 19.02g were caged individually in quiet laboratory at room temperature with 12 hour light dark cycle. Mice have free access to water and food (ad libitum). The wash out of the drugs administered is considered and after each dose the tested animals were given the break of 1 day.

Agents:
Control group was on rodent feed however Reference group was treated with alprazolam dose (0.5mg/60kg). The 2 test group received ficus carica fruit dilution at the doses of 250mg/kg/100ml & 500mg/kg/100ml for their anxiolytic potentials were based on preliminary evidence of ethanolic extract of ficus carica fruit 100 and 200mg/kg body weight of rats being use as a diuretic agent. Experimental Protocol: The mice were first taken into elevated plus maze and then into light/dark activity box to induce anxiety and fear in them. The anxiety and activity behavior were noted as control. The forced swim test (FST) is a preclinical test to the screening of antidepressants. The mice were induced depression by this test and then their behavior changes were examined by using hole-board apparatus. After this the mice of each group were given the test and reference drug in order to discover the changes in their mental activity i.e.
anxiolytic and antidepressant action of the drugs.

Anxiolytic screening by Using Light/Dark Box Method
Anxiety and activity corresponding variables in mice were evaluated by submitting the mice in light/dark box for 10min after 40min and 24hours of administration for both test groups. The activity was compared with alprazolam reference dose of 0.5mg/60kg. Mice were submitted for 10min in light/dark box one by one after 40min of drug administration. Anxiolytic screening by using Elevated plus Maze Elevated plus maze is an excellent model for determination of antidepressant and anxiolytic effect in mice. It is 50cm above from the base and 10cm wide. The mice were placed in center of the all four arms with its face towards open arm. The behaviors of each group of mice were noted for 5 minutes. The test group mice were treated with two different dilutions of fig and were analyzed one by one after 40min and 24hours. The activity was compared with alprazolam reference dose. Mice were submitted for 10min in light/dark box one by one after 40min of drug administration.

Antidepressant screening by using Head Dip Method/ Hole-Board Apparatus
This Hole Board apparatus is used to evaluate antidepressant effect in mice after inducing depression by the aid of force swim method. Variation in the psychological state of mice were examined by observing changes in exploratory activity, i.e. numbers of head-dip after depression being induced in mice and then when the reference and fig dose are provided. The numbers of head dips by mice were observed 40min and 24hours post administration of the test doses respectively and 40min post reference dose .The mice were submitted to hole-board apparatus for 5min.

Result
The experimental data obtained from light/ dark box test, head dip method and elevated plus maze test clearly reveals antidepressant & anxiolytic activity of ficus carica fruit. The results showed that on lower doses 250mg/kg/100ml the antidepressant and anxiolytic action of the fruit appears to be significant and by increasing the dose to 500mg/ kg/100ml, marked and pronounce anxiolytic and antidepressant effects were noted in mice. This effect was comparable with a well-known drug alprazolam shown in Table 1 Figure 1]. The control group mice hesitate to explore the field and remain confined to the close chamber upon application of both the method to check anxiety and anxiolytic effects but mice of reference and test groups showed great interest to explore both the fields [ Table 2 & Figure 2].  Figure 3].  500mg stayed most of the time in open field (anxiety index) even more than the mice of reference group. The mice of these two groups have alleviated level of tension and stress. The mice spend time in close arm of test group 2 and reference group are far more shorten than the control group. Table 3 demonstrates the antidepressant activity of mice by using head dip method. After performing force swim test, the mice were induced depression and they showed greater latency but when the test doses and fig dose were provided the mice showed improved motor activity. By observing these tables we can say that the high amount of polyphenols present in fig bears powerful action against fear, anxiety and depression in dose of 500mg/kg/100ml of water. It claims to have more antidepressant & anxiolytic action than a widely prescribed drug alprazolam. It has been reveal from this study that a slight higher dose of fig could improve its anxiolytics well as antidepressant capabilities.

Conclusion
Ficus carica linn as a dried fruit easily available & have remarkable source of nutrients, proven to have active anxiolytic and antidepressant activity. Since this study was purely confined to the mice, there's much probability and possibilities to have similar effects in humans too but it will required further research.