Evaluation of Antidepressant-like Effect of Clitoria ternatea Linn.

 

Shruti Mittal1*, Prashant Gupta1, Vijay Nigam2

1Department of Pharmacology, Daksh Institute of Pharmaceutical Science, Chhatarpur, India.

2Department of Pharmacognosy, Daksh Institute of Pharmaceutical Science, Chhatarpur, India.

*Corresponding Author E-mail: shrutimittal2912@gmail.com

 

ABSTRACT:

Depressive disorder is a prevalent psychiatric disorder, which affects 21% of the world population. The presently using drugs can impose a variety of side-effects including cardiac toxicity, hypopiesia, sexual dysfunction, body weight gain, and sleep disorder. Ayurvedic medicine may be a powerful weapon given by our nature to cure disease. Considering the importance of plants as sources of drugs even today people are adopting different herbal drugs for the treatment of assorted diseases. During the last decade, there is a growing interest in the therapeutic effects of natural products on mental disorders. This study planned to assess antidepressant like activity of methanolic extract of Clitoria ternatea Linn. (fabaceae). Soxhlet extraction method was used for methanolic extraction. Antidepressant activity was studied using forced swimming test (FST) and tail suspension test (TST). Two doses 200 and 400 mg/kg of methanolic extract of flower were selected for testing. Imipramine (10 mg/kg, i.p.) were used as the reference standard drugs. Methanolic extract of Clitoria ternatea flower significantly reduced immobility time in both TST and FST. Extract increased the climbing behavior in FST, which is similar to effect observed with imipramine. The results of this study suggest that antidepressant like effect of Clitoria ternatea seems to be mediated by an increase in norepinephrine level in synapses. However further study is needed to understand mechanism of action and to isolate the active component responsible for antidepressant like activity.

 

KEYWORDS: Antidepressant-like effect, Clitoria ternatea, Forced swimming test, Tail suspension test, Depression.

 

 


INTRODUCTION:

Mood disorders are common in psychiatry and their characteristic is disturbance in regulating emotions. People affected with mood disorders, experience different emotions ranging from irritability to chronic depression. 1 Depression in late life is associated with significant morbidity, including deficits in a range of cognitive functions and considerable influence on functional impairment, disability, decreased quality of life and has a negative effect on the body’s recovery from illness, increases the rate of suicide, increases use of health care services and expenses and can result in early death and disturbance in the general state of wellness. 2

 

The exact cause of depression is not known. Many researchers believe it is caused by chemical changes in the brain. This may be due to a problem with your genes, or triggered by certain stressful events. More likely, it's a combination of both. 3 There are several types of depression. A person with major depression experiences symptoms of depression that last for more than two weeks. A person with dysthymia experiences episodes of depression that alternate with periods of feeling normal. A person with bipolar disorder, manic-depressive illness, experiences recurrent episodes of depression and extreme elation. A person with Seasonal Affective Disorder (SAD) experiences depression during the winter months, when day length is short. Although the exact cause of depression is unknown, research suggests that depression is linked to an imbalance of the neurotransmitters serotonin, norepinephrine, and dopamine in the brain. Factors that may contribute to depression include heredity, stress, chronic illnesses, certain personality traits (such as low self-esteem), and hormonal changes. 4 Depression is a significant contributor to the global burden of disease and affects people in all communities across the world. Today, depression is estimated to affect 350 million people. The World Mental Health Survey conducted in 17 countries found that on average about 1 in 20 people reported having an episode of depression in the previous year. 5 By 2020 it is expected to be the second-leading cause of disability globally. 6 At its worst, depression can lead to suicide, a tragic fatality associated with the loss of about 850 000 lives every year. 7 Animal model (Tail suspension test and Despair swim test) were used for evaluating antidepressant activity. There have many Ayurvedic plants/medicine which are effectively used for the psychiatric disorders.

 

Ayurveda, being a stream of treatment, can no way ignore the importance of psychic condition of a patient to effectively diagnosis and treat disorders. Medical science admits that the root cause of multiple disorders lies in psychic tensions. Ayurveda has a well-developed branch of psychiatry, which has clinically proven practices of treating mental ailments and also physical problems associated with psychological imbalances. 8

 

Ayurveda the traditional system of Indian medicine mentions a number of plant products which can be used in the treatment of psychiatric disorders. The Ayurvedic concept of Rasayana consists of specialized class of drugs which prevent ageing, increase longevity, impact immunity, improve mental functions and vitality to body a number of synthetic drugs are being used as standard treatment for clinically depressed patient, they have adverse effects that can compromise the therapeutic treatment. Thus, it is worthwhile to look for antidepressant from plants with proven advantage and favorable benefit to risk ratio. A number of medicinal plants and medicine derived from these plants have shown antidepressant properties by virtue of combined effect of their medicinal constituents. 9 Synthetic antidepressants are often associated with their anticipated side effects like dry mouth, inability in driving skills, constipation and sexual dysfunction and majority of patients are reluctant to take this treatment. 10

 

Clitoria ternatea L. belongs to family Fabaceae and it is commonly known as ‘Butterfly pea’. It is a perennial twining herb, found throughout India in tropical areas. [11]Clitoral ternatea comprise 60 species distributed mostly within the tropical belt.[12]Flower is an important part of plant which contains a great variety of natural antioxidants, such as phenolic acids, flavonoids, anthocyanin and many other phenolic compounds. Flower part of plant contain excessive amount of Tannin. [13] The present study was aimed to perform a systematic study and to investigate the possible antidepressant activities of methanolic extracts of clitoria ternatea in normal and in experimentally induced depressant animal. Plant show a number of pharmacological effect that include- Antimicrobial effect, Anti-inflammatory antipyretic and analgesic effects, Antidiabetic Effect Wound healing effect, Hepatoprotective effect, Antidiarrheal activity, Immuno-modulator activity, Effects in CNS, Local anesthetic activity. Anticancer activity, Diuretic activity, Gastrointestinal effect, Anti Asthmatic activity, Hypolipidemic effect.[14]

 

MATERIAL AND METHOD:

Drugs:

Drug (Imipramine), chemicals, instruments, equipment’s used in this study were procured from labs of Daksh Institute of Pharmaceutical Science, Chhatarpur (M.P.). All the other solvents and chemicals used are of analytical grade.

 

Plant material:

Flowers of Clitoria ternatea plant were collected from forest area of Bundelkhand region, distt- Chhatarpur (M.P.). They were authenticated by Dr. Pushpendra Kumar Khare, Assistant professor of Botany, Govt. Maharaja PG College, Chhatarpur (M.P.) and were given a specimen no. (041/Bot/2019).

 

Preparation of Extract:

The flowers of plant were shade dried and then powdered in a mechanical grinder for preparation of extract. The powdered materials of flowers were Soxhlet extracted with 90% methanol for about 8 hr. The extract was concentrated by distilling the solvent to obtain the crude extract. Finally, the extract was evaporated to dryness. The extract, on removal of solvent in vacuum, gave a dark greenish brown semisolid residue. The powdered material or the extracts of the flowers are used for the experimental procedure.

 

Experimental Animals:

Albino mice (weight 20–25 g) and Albino rats (160-200 g) of either sex are used. Animals were housed under standard conditions of temperature (24±20C) and relative humidity (30-70%) with a 12:12 (light: dark) cycle. The animals were given standard diet and water at libitum. All procedures and protocols were approved by Institutional Animal Ethics Committee(Protocol no. DIPS/IAEC/NO: 09/2018-19).

 

Experimental Group

Group-I treated as Vehicle. (Water)

Group-II treated as Standard drug (Imipramine 10mg/kg)

Group-III treated as Test drug (200 mg/kg)

Group-IV treated as Test drug (400 mg/kg)

 

METHODS:

Tail Suspension Test:

Animals are transported from the housing room to the testing area in their own cages and allowed to adapt to the new environment for 1 h before testing. They are brought to the laboratory at least one day before the experiment and are housed separately in cages with free access to food and water.4 Group of 6 animals are treated with by intra-peritoneal injection 30 minutes prior to testing. For the test, the mice were suspended on the edge of a shelf about 58 cm above top of table by adhesive tape and placed approximately 1cm from the tip of the tail. The duration of immobility is recorded for periods of 5 min. Mice are considered immobile when they hang passively and completely motionless for at least 1 min.15

 

Despair Swim Test:

Naive rats are individually forced to swim inside a vertical Plexiglas cylinder (height: 40 cm; diameter: 18 cm, containing 15 cm of water maintained at 25 °C). Animals placed in the cylinders.  After placing for the first time, animals were highly active initially and they vigorously swimming in circle then they trying to climb the wall or diving to the bottom. Activity begins to fall down and to be interspersed with phases of immobility or floating of increasing length, after 2–3 min of experiment. After 5–6 min, immobility in rats reaches a plateau where the rats remain immobile for approximately 80% of the time. After 15 min in the water the rats were removed and allowed to dry in a heated enclosure (32 °C) before being returned to their home cages. They are again placed in the cylinder after 24 h later of previous experiment and the total duration of immobility is measured during a 5 min test. Floating behavior during in this 5 min period has been found to be reproducible in different groups of rats. Whenever animals remains floating passively in the water in a slightly hunched but upright position, its nose just above to the surface, an animal is judged to be immobile. 4 Group of 6 animals are treated with by intra-peritoneal injection. Test drugs or standard were administered to animals prior one hour to testing. Since experiments with the standard drug (Imipramine) showed that injections 1, 5 and 24 h prior the test gave the most stable results in reducing floating these times are chosen for the experiment.16

 

Statistical Analysis:

Data were analyzed by Graph Pad Prism version 8.1.1 software and presented as mean and SEM values. The statistical tests used one-way analysis of variance (ANOVA) followed by Dunnett’s multiple comparison test.

 

RESULT:

Table:1 Percentage yield of fresh weight and dried weight of Clitoria ternatea Linn flowers.

 

Plant

Weight (gm)

Dry weight (gm)

Percentage yield

Clitoria ternatea Linn.

35

6.03

17.22

 

Table:2 Qualitative analysis of the methanolic extracts of Clitoria ternatea to screen for the   presence of phytochemicals.

S. No.

Phytochemicals

Plant parts

Flower

1.

Alkaloids

+ve

2.

Flavonoids

+ve

3.

Tannin

+ve

4.

Glycoside

+ve

5.

Resin

+ve

6.

Steroids

-ve

7.

Saponin

-ve

8.

Phenol

-ve

+ Presence of the compound.

- Absence of the compound.

 

For the Qualitative analysis, we analyzed the methanolic extract of Clitoria ternatea flower. We analyzed that, this extract confirmed the presence of flavonoids, tannins, alkaloids, glycosides, resin. Then we have done the Quantitative analysis.

 

Table. 3 Quantitative analysis of the methanolic extracts of Clitoria ternatea for estimation of phytochemicals.

Phytochemicals

Plant parts

Average Estimated Value (mg/gm)

Flavonoids

Flowers

42±0

 

When we go for the Quantitative analysis for estimation of phytochemicals, we found that, flower extract content 42 mg/gm of flavonoids.

 

Pharmacological Analysis:

The antidepressant effects of Clitoria ternatea (200 and 400 mg/kg) and Imipramine were studied by observing the changes in the duration of immobility in the two models, Forced swim test (FST) and Tail suspension test (TST).

 

Tail suspension test:

Table: 4 Effect of methanolic flower extract of Clitoria ternatea on duration of immobility time in the Tail suspension test

S. no

Groups

Treatment

Tail suspension test

Duration of immobility(in sec)

1.

Control group

Vehicle (water)

160.2±2.45

2.

Test group

Clitoria ternatea(200 mg/kg)

69.3±1.6**

3.

Test group

Clitoria ternatea (400 mg/kg)

74.2±1.26**

4.

Standard group

Imipramine (10 mg/kg)

62.1±1.07***

Values represented as Mean ± SEM  **P< 0.05 vs control group for test solutions and for standard group represent the significant value***P<0.001vs control group.

 

Forced swim test-

 

Table:5 Effect of methanolic flower extract of Clitoria ternatea on duration of immobility time in the Forced swim test

S. no

Groups

Treatment

Forced swim test

Duration of immobility (in sec)

1.

Control group

Vehicle (water)

140±1.11

2.

Test group

Clitoria ternatea (200 mg/kg)

51.8±1.59**

3.

Test group

Clitoria ternatea (400 mg/kg)

59.2±0.76**

4.

Standard group

Imipramine (10 mg/kg)

61.6±0.58***

Values represented as Mean ± SEM  **P< 0.05 vs control group for test solutions and for standard group represent the significant value***P<0.001vs control group.

 

DISCUSSION:

A number of side effects like liver damage, cardiac problems and mutagenesis are associated with the use of antianxiety drugs obtained from synthetic sources. Thus, researchers are exploring natural resources to find out newer and safer natural anxiolytic agents.

 

We have collected about 50 gm of fresh flower of Clitoria ternatea, after shaded drying, we got 35 gm of flowers. Then, flowers subjected for the extraction process by Soxhlet extraction method by using methanol as a solvent. After extraction, we got 6.03 gm of methanolic extract and got % yield of methanolic extract 17.22 %. Preliminary phytochemical studies showed presence of flavonoids and triterpenoids in methanol extract of C. ternatea. The results of present investigations are in agreement with the available literature where flavonoids – kaempferol17 and apigenin[18] and triterpenoids – α, β-amyrin have been reported to exhibit antianxiety activity. Active compounds found in various pieces of plants have been appeared to have remedial qualities. Utilizing entire plant as a medication would thus be able to be utilized to receive greatest helpful rewards.[19] Flavonoids found in various medicinal plants have been appeared to have antianxiety and depression reducing impact, for example, Hypericum perforatum.20

 

Anxiolytic activity of methanol extract of blue variety of C. ternatea flower was evaluated employing widely used model, i.e., FST and TSM. The FST model was chosen since these are effective, cheap, simple, less time consuming and do not cause much discomfort to the animals while handling18, 21. The models are principally based on the observations that exposure of animals to approach–avoidance conflict which is manifested as an exploratory-cum-fear drive.

 

In TST, Clitoria ternatea 200 and 400 mg/kg, (intra peritoneal) produced significant reduction (p<0.05 respectively) in the immobility period when compared with that of control group animals that received only the vehicle. The extract (400 mg/kg) was found to be effective and it exhibited activity similar to that of the standard drug Imipramine (p<0.001). In FST, Clitoria ternatea 200 and 400 mg/kg, (intra peritoneal) produced significant reduction (p<0.05 respectively) in the immobility period when compared with that of control group animals that received only the vehicle. The extract (400 mg/kg) was found to be effective and it exhibited activity similar to that of the standard drug Imipramine (p<0.001).The above results indicated that methanolic extract of C. ternatea possess anti anxiety activities. The CNS depressant activity was also observed at higher dose levels. The traditional claim of C.ternatea being used as anxiolytic has been scientifically validated.

 

It is suggested that methanol extract of C. ternatea may be act via binding to benzodiazepine receptors as agonist22 increase ascorbic acid level in brain23, inhibition of γ-amino butyric acid transmission24 and monoamine oxidase inhibition. Further, it is concluded that triterpenoids or flavonoids may be responsible for antianxiety activity of both varieties of C. ternatea. It can be finally concluded that these phytoconstituent (s) may be isolated using column chromatography to develop anxiolytic agent.

Since ancient times, people have been using a no. of plants in various ways as a source of medicine. From the above preclinical study, we can conclude that methanolic flower extracts of Clitoria ternatea show a significant antidepressant activity in TST and FST models of depression. We believe that Clitoria ternatea has the potential to be used as an adjuvant in the treatment of depression and other mood disorders. To gain closer insights into the exact mechanism of its action, further research is required.

 

ACKNOWLEDGEMENT:

 The financial support provided by the Daksh Institute of Pharmaceutical Sciences, Chhatarpur (M.P.) for the present research work is duly acknowledged.

 

CONFLICT OF INTEREST STATEMENT:

We declare that we have no conflict of interest.

 

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Received on 10.08.2020            Modified on 15.10.2020

Accepted on 26.11.2020           © RJPT All right reserved

Research J. Pharm.and Tech 2021; 14(12):6437-6441.

DOI: 10.52711/0974-360X.2021.01113