INTRODUCTION:

Depression is a common mental disorder that presents with depressed mood, loss of interest or pleasure, decreased energy, feelings of guilt or low self-worth, disturbed sleep or appetite, and poor concentration¹. Depression has a significant impact on an individual’s ability to perform life activities². People who were born in the later part of the 20th century seemed to have higher rates of depression and suicide than those of the previous generation, in part, because of high substance abuse and the rising demands in the standards of living^{3, 4}. Anti-depressant drugs which are used in the Allopathic system of medicine for the treatment of depression have side effects⁵.

Nature always stands as a golden mark to exemplify the outstanding phenomena of symbiosis⁶. One of the advantages of herbal medicines is their complex composition. Their components have multiple activities that result in a greater total activity⁷.The phytochemical profile of this plant Chrozophora plicata reveals the presence of flavonoids, triterpenoids, saponins and alkaloids. It is evident from the literature that flavonoids⁸and triterpenoids⁹posseses antidepressant properties. Since there are no reports of isolation of active antidepressant principles of Chrozophora plicata, an herbal ingredient present in unani medicine Safi, the present study is planned to exploit the mood elevating activity of herbal plant named Chrozophora plicata, family: euphorbiaceae by using tail suspension test and despair swim test in albino mice.

MATERIALS AND METHODS:

For the present study, the leaves of Chrozophora plicata were collected from the surrounding gardens of the Gajwel (Mandal), Medak Dist, Andhra Pradesh, India. After the fresh leaves were authenticated by national bureau of plant genetic resources, Rajendranagar, Hyderabad, leaf specimens have been deposited at the museum of the college. Fresh mature leaves were shade dried at room temperature, coarse powdered and extracted with petroleum ether by soxhlet’s extraction method. Thereafter, the extracts were concentrated using electric water bath to obtain semisolids crude extract. The percentage yields of the leaf extract were found to be 10.6%. The extract was stored in airtight container in refrigerator below 10ºc. Appropriate concentration of stock solution of extract were prepared using 2% acacia suspension and used for the following studies.

Preliminary phytochemical screening¹⁰

Preliminary phytochemical tests were performed for the petroleum ether extract of Chrozophora plicata to detect the presence of phytochemicals by following the standard methods described in the Practical Pharmacognosy of Kokate and Khandelwal. The results have been tabulated in table I.

Experimental animals:

Albino Mice (18-25g) were used in the experiment. They were procured from Sainath Agencies, Musheerabad. After randomization into various groups and before initiation of experiment, the mice were acclimatized for a period of 10 days. Animals were housed in polypropylene cages and maintained under standard environmental conditions such as temperature (26 ± 2ºc), relative humidity (45-55%) and 12hr dark/light cycle. The animals were fed with rodent pellet diet (Golden Mohur Lipton India Ltd.) and water ad libitum. The study protocol was approved from the institutional animal ethics committee (IAEC) before commencement of experiment (1230/a/08/CPCSEA).

Determination of acute toxicity:

The Chrozophora plicata petroleum ether extract was studied for acute toxicity study at a dose of 5 mg/kg, 50mg/kg, 300 mg/kg, and 2000 mg/kg p.o in albino mice. The extract was found safe to all the animals 12 mice. The mice are subjected to a dose of 5000 mg/kg. Even at 5000mg/kg no mortality is seen, but few symptoms of CNS depression such as sedation, drowsiness and motor in coordination is seen in all the mice at 5000mg/kg. Hence a dose of 3000mg/kg is selected as safer dose and 1/5^th of 3000mg/kg i.e. 600mg/kg is selected for our study.

Evaluation of antidepressant activity of petroleum ether extract of Chrozophora plicata leaves by tail suspension test in albino mice:

The experiment was performed on albino mice (18-25gms) of either sex procured from Sainath Agencies, Musheerabad. The animals were housed in colony cages at an ambient temperature of 26+2⁰C and, relative humidity (45-55%),with a 12h/12h light dark cycle and access to food and water ad libitum. Food was restricted during experiments. Stock solutions of amitriptyline (20mg/kg) and petroleum ether extract of Chrozophora plicata leaves (600mg/kg) were prepared in 2% acacia suspension. Weigh and mark the animals. Divide the animals in to three groups control(C), test (T), and standard(S) each consisting of 3 mice. The control group mice were suspended by their tail using 75 cm long thread for a period of 6mins and total duration of immobility was recorded. Repeat the same procedure for test group and standard group mice after 1hr of administration of petroleum ether extract of Chrozophora plicata leaves and amitriptyline (20mg/kg, p.o) and their immobility time was recorded. A mouse is considered as immobile when it gets freezed without performing any muscular activity.

Evaluation of mood elevating activity of petroleum ether extract of Chrozophora plicata leaves by using despair swim test in albino mice:

Each animal of control group is placed in a transparent beaker (25cm height × 10 cm diameter) filled to 6cms depth with water at room temperature. The study was initiated, which consisted of allowing the animals to swim for 10mins. The duration of immobility was recorded during this period. A mouse was judge to be immobile when it floated in an upright position, making only small movements to keep its head above water. Repeat the same procedure for test group after 1hr of administration of petroleum ether extract of Chrozophora plicata leaves(600mg/kg) and for standard group after 1 hour administration of amitriptyline (20mg/Kg p.o),and their immobility time was recorded.

Following both the swim sessions the animals were removed from beaker, dried with towels and placed in a heated cage for 15mins before being return to their cages.

Statistical Analysis:

The values are represented as mean ± S.E.M, and statistical significance between treated and control groups was analyzed using One way ANOVA, Followed by students t test where P<0.001, P<0.01 and P<0.05 was considered statistically significant.

RESULTS AND DISCUSSION:

Preliminary Phytochemical screening of petroleum ether extract of Chrozophora plicata leaves reveals the presence of flavonoids, triterpenoids, saponins and traces of tannins. The results are shown in table I. In tail suspension test induced depression model it is observed that the duration of immobility of Chrozophora plicata (600mg/kg) treated mice (80.6 ±1.51seconds) is less than the duration of immobility observed with control treated (distilled water, p.o) mice (131.3 ±0.62seconds). The results are shown in tables II, III and IV which is a clear indication of inhibition of depression with petroleum ether of Chrozophora plicata leaves. In forced swimming test depression model it is observed that the control treated mice fails to make vigorous attempts to swim in a beaker exhibiting duration of immobility of 295± 1.47 seconds. Whereas the extract treated mice makes more vigorous attempts to swim in a beaker showing less duration of immobility of 245.6± 0.05 seconds. The results are depicted in tables V, VI andVII. Chrozophora plicata leaves are available abundantly throughout the Andhra Pradesh and is an essential ingredient of unani herbal medicine Safi. The exact mechanism behind antidepressant activity of Chrozophora plicata is not understood. But it could be assumed that Chrozophora plicata leaves (600mg/kg) posses mood elevating activity by enhancement of serotonin and norepinephrine levels in synapses of brain neurons. However the exact biochemical mechanism of Chrozophora plicata leaves in alleviating depression in experimental mice is yet to be established.

Table I: Preliminary Phytochemical screening of Petroleum ether extract of Chrozophora plicata leaves.

Phytoconstituents

Petroleum ether leaf extract

Flavonoids

Triterpenoids

Saponins

Alkaloids

Tannins

Glycosides

+++

- Absent

++ Present

+ ++ Present with more clarity

Tables II, III and IV representing antidepressant activity of Chrozophora plicata leaves by using tail suspension test in mice.

Table II: Control: (Distilled water p.o)

S.No

Body Weight (gms)

Duration of Immobility(Sec)

145±0.11

120±0.32

129±1.47

Mean =131.3 ±0.62sec

Table III: Test (Pet. ether extract of Chrozophora plicata 600mg/kg, p.o)

S. No	Body weight (gms)	Volume of drug to be administered in ml	Duration of immobility (Sec)
1	24	0.24	65±0.15***
2	22	0.22	90±1.71
3	19	0.19	87±1.39**
			Mean=80.6**±1.51sec

Table IV: Standard Group :(Amitriptyline 20mg/Kg)

S.NO

Body Weights

(gms)

Volume of Drug To Be Administered (ml)

Duration Of Immobility(Sec)

0.24

0.18

0.20

107±1.54

37±0.28

75±0.03***

Mean=73±1.02***sec

P<0.001***, P<0.01** and P<0.05* was considered statistically significant.

Tables V, VI and VII represents antidepressant activity by using forced swimming test in mice.

Table V: Control (distilled water, p.o)

S. No	Body weight (gms)	Duration of immobility (Sec)
1	21	350± 0.92
2	18	330±1.58
3	24	206±1.01

Mean= 295± 1.47seconds

Table VI: Test Group (Petroleum ether extract of Chrozophora plicata 600mg/kg leaf extract, p.o)

S. No.	Body Weight (gms)	Volume of drug to be administered (ml)	Duration of immobility (seconds)
1	24	0.24	255±1.87*
2	19	0.19	240±0.53
3	26	0.26	242±0.14

Mean= 245.6± 0.05*seconds

Table No.VII: Standard Group: (Amitriptyline 20mg/kg, p.o)

S. No.	Body Weight (gms)	Volume of drug to be administered (ml)	Duration of immobility (Sec)
1	25	0.25	104±0.64***
2	20	0.20	240±1.30*
3	24	0.24	122±1.68*

Mean =155.3±1.95**sec

P<0.001***, P<0.01** and P<0.05* was considered statistically significant.

Fig I: Antidepressant activity of Chrozophora plicata leaves by using Tail suspension test in mice.

Fig II: Antidepressant activity of Chrozophora plicata leaves by using Forced swimming test in mice.

Fgure III

Figure IV

Fig III and IV: Graphical representation of the results obtained from Antidepressant activity of methanolic extract of chrozophora plicata leaves (600mg/kg) by using tail suspension and forced swimming test in mice.

CONCLUSION:

The data obtained from the present study indicates that the petroleum ether extract of Chrozophora plicata leaves at 600mg/kg possessed significant mood elevating effect and thus supports the use of Chrozophora plicata leaves in treatment of major depression. Tail suspension test and despair swim test models causes depression by increasing pessimism and distress in mice when subjected to depression. However petroleum ether extract of Chrozophora plicata leaves had significantly reduced pessimism and desperation in mice indicated by decrease in the duration of immobility in mice. Hence the data obtained from the present research on petroleum ether extract of Chrozophora plicata leaves suggests the use of the plant in major depression.

ACKNOWLEDGEMENTS:

The authors are grateful to management of Vijaya College of Pharmacy, Hyderabad for providing the facilities for our Research.

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Received on 19.02.2014 Modified on 19.03.2014

Res. J. Pharmacology & P’dynamics. 6(2): April- June 2014; Page 75-78