Anti-inflammatory Activity of Aqueous and DCM Extracts of Jatropha gossypifolia Linn in Albino Rats

Akshada Kakade*, Indrayani Raut, Sandeep Kane, Rahul Adnaik, C. S. Magdum, and S. K. Mohite

Rajarambapu College of Pharmacy, Kasegaon, Maharashtra.

ABSTRACT:

Medicinal herbs are the local heritage with global importance. World is endowed with a rich wealth of medicinal herbs. The different variety of plants with different therapeutic properties is quiet astonishing. Jatropha gossypifolia Linn (Euphorbiaceae) is a large genus of shrubs, herbs and trees distributed in the tropical parts of the world. A decoction of the bark is used as an emmenagogue, that of the leaves for stomach ache, venereal diseases and as blood purifier. The plant has been used ethnomedically to prevent cancerous growth. The plant is very popular in all Maharashtra and Jatropha gossypifolia Linn has suggested in the Indian system of medicine for number of diseases. The plant is known to possess various active agents such as flavonoids, triterpenoids, sterols (phytosterol), tannins, essential and volatile oils. The anti inflammatory activity of the aqueous and dichloromethane extract of the leaves of Jatropha gossypifolia Linn was investigated with experimental animal model using the carrageenan induced rat paw edema method. The extract 200mg/kg at 240min post treatment caused a significant (p<0.05) reduction in the paw oedema in rats. The effect of the extract was more pronounced at the dose 200mg/kg and was closer to that of ibuprofen 10mg/kg. The finding of the studies indicate that the aqueous extract and dichloromethane extract of leaves of Jatropha gossypifolia Linn had good anti-inflammatory activity in carrageenan induced hind paw edema in rats between 2 to 3 hours.

KEYWORDS: Jatropha gossypifolia Linn, Euphorbiaceae, Anti-inflammatory activity, carrageenan, paw edema.

INTRODUCTION:

The dried stem bark of the plant contains an intensely bitter amorphous alkaloid, jatrophine which is similar to quinine in properties. The bark also contains resins, isophytosterol and tannin. The latex is poisonous and contains 2.5% alcohol soluble matter.

Tender leaves contain a pentose glycoside of cyanidine. The seeds contain fatty acids, β-sitosterol, amino acids, long chain alcohols. The leaves and the seeds are used as purgatives⁵. The plant has been used ethno medically to prevent cancerous growth⁶.

The plant has excited a considerable amount of interest because of its important medicinal activity, novel and complex metabolites. The plant is very popular in all Maharashtra and Jatropha gossypifolia Linn has suggested in the Indian system of medicine for number of diseases. The plant is known to possess various medicinal properties.

The medicinal plants might provide a useful source of new active compounds for development of new pharmaceutical entities. Screening of these compounds gives birth to new pharmacological activity².

Literature survey reveals that no systematic approach has been made to study the anti-inflammatory activity of leaves of this plant. In the present work, we have investigated the anti-inflammatory activity of the aqueous and dichloromethane extract of Jatropha gossypifolia Linn using animal model.

Materials and Methods^11-14:

The fresh leaves of Jatropha gossypifolia Linn were collected in the month of July from the local area of Sangli region, Maharashtra. The plant was authenticated by Dr. A. K. Madgum, H.O.D of Botany Department, Willingdon College, Sangli.

The dried material was crushed using mechanical grinder. The coarse powder of leaves of Jatropha gossypifolia Linn weighing 60 gm was taken in a thimble of filter paper. The diameter of thimble was little lesser than the inner diameter of soxhlet apparatus.

The successive extraction was carried out³. This was followed by using polar to non polar solvents. An aqueous extract was also prepared by maceration method using chloroform water I.P. for seven days with occasional shaking. The dichloromethane and aqueous extract was concentrated to dryness under reduced pressure and controlled temperature of 60⁰C to yield solid masses that are completely free from solvents⁷. The pharmacological screening was carried out using standard protocols.

Animals

Healthy, albino rats (Wistar strain) weighing from 150 – 165 gms were selected for present investigation. The animals were housed individually in a room maintained under environmentally controlled conditions of 24 ± 1º C and 12 hr light – 12 hr dark cycle with free access to food and water during the course of experiments. Animals were fed with standard laboratory diet (SLD).

Acute toxicity study⁸

Acute toxicity study was carried out according to OECD guidelines. Swiss albino mice (wt-20-25 mg) were administered oral dose (1000 mg/kg, 1200 mg/kg, 1400 mg/kg, 1600 mg/kg, 1800 mg/kg, 2000 mg/kg.) of aqueous and dichloromethane extract of leaves of Jatropha gossypifolia Linn. Each dose group contains five mice. After administration of dose of extract the animals were observed for toxic effect after 24 hours. The toxicological effect was observed in terms of mortality. Observation of mice after 24 hours showed no mortality. The final dose decided was 200 mg/kg for the experiment.

Carrageenan induced rat paw edema method^9-10:

Fig no 1 Plethysmometer

Wistar rats (150-165 gms) of either sex were maintained under standard environmental conditions. They had free access to standard diet and water. Anti-inflammatory activity was measured using carrageenan induced rat paw edema method. First group of five rats were given a dose of dichloromethane extract (200 mg/kg) of leaves of Jatropha gossypifolia Linn. Second group were given a dose of 200 mg/kg of aqueous extract of leaves of Jatropha gossypifolia Linn. After 1hour, 0.1 ml of 1% carrageenan suspension in 0.9% normal saline solution was injected into the sub-plantar tissue of right hind paw. The volume displaced by hind paw was measured at interval of one hour with plethysmometer for next four hours. Standard (Ibuprofen) in a dose of 10 mg/kg was given to third group. Fourth group was maintained as a control. The mean paw edema value for the test group was compared with its mean value for the control group. Reduction in the edema volume was taken as a measure of anti-inflammatory activity. T he effect of the extract was more pronounced at the dose 200mg/kg and was closer to that of ibuprofen 10mg/kg. Percentage (%) edema inhibition was also calculated for comparison.

Table No. 01 Showing mean paw volume in ml for anti inflammatory activity

Groups	Mean paw volume in ml
Groups	0 Hr.	1 Hr.	2 Hr.	3 Hr.	4 Hr.
Control (N.S)	0.0833 + 0.1856	0.4366 + 0.2478	2.6966 + 0.589	2.9133 + 0.3175	2.633 + 0.2876
Standard (Ibuprofen) (10 mg/kg)	0.066 + 0.012	0.2966 + 0.0949	0.6966 + 0.2976	0.7466 + 0.2820**	0.3033 + 0.0783*
Aqueous extract (200 mg/kg)	0.07663 + 0.0033	0.34 +0.0808	0.9633 + 0.1041*	0.9133 + 0.4199**	0.8433 + 0.3968*
Dichloromethane extract (200 mg/kg)	0.07 + 0.0057	0.3566 + 0.1342	0.92 + 0.7206	0.9866 + 0.7130**	0.92 + 0.711

Values are expressed as mean + SEM (n=5)

** P<0.05 compared with control, considered significant, (ANOVA followed by Dunnett’s t-test)

Table No 2 Showing % edema inhibition for anti-inflammatory activity

Group	% edema inhibition
Group	0 Hr.	1 Hr.	2 Hr.	3 Hr.	4 Hr.
Aqueous extract (200 mg/kg)	8.75%	22.01%	74.40%	68.76%	68.09%
Dichloromethane extract (200 mg/kg)	15.66%	19.72%	65.87%	66.35%	65.05%
Standard (Ibuprofen) (10 mg/ml)	27.71%	33.48%	74.40%	74.59%	88.60%

Statistical analysis:

The data was analyzed using one way analysis of variance. ANOVA using Dunnett’s t-test were carried out for the analysis to determine significant overall effects (P<0.05).

Results:

The present phytochemical investigation on Jatropha gossypifolia Linn leaves reveals the presence of terpenoids in dichloromethane and aqueous extract. Hence we have taken these extracts for evaluation of anti-inflammatory activity.

Carrageenan induced rat paw edema method:

This anti inflammatory activity was dose dependent and found to be statistically significant at the higher concentration, 200mg/kg, Table no 01. % edema inhibition for extracts shows good comparative results with that of standard percentage values, Table no 02. The anti inflammatory activity of ibuprofen, a standard reference drug was also found to be significant.

DISCUSSION:

The crude extract showed presence of multiple chemical constituents with presence of flavoinds and terpenoids. The dichloromethane and aqueous extract are devoid of toxicity in albino rats. This anti inflammatory activity was found to be dose dependent and statistically significant. However, this activity was less potent as compared to ibuprofen. The anti inflammatory activity of Jatropha gossypifolia Linn appears due to significant reduction of various biochemicals viz histamine, 5HT, various kinins which are involved in early phase of inflammation. Activity is due to synergistic effects of various phytoconstituents present in it. At this stage, it is difficult to say that which exact constituents is responsible for activity. However, further active phytoconstituents isolation is needed to carry out the activity.

ACKNOWLEDGEMENT:

We are thankful to the Principal of Rajarambapu College of Pharmacy, Kasegaon and Appasaheb Birnale college of Pharmacy, Sangli for providing necessary facilities for carrying out research work.

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Received on 31.12.2010

Accepted on 27.02.2011

Research Journal of Pharmacognosy and Phytochemistry. 3(4): July- August 2011, 148-150