Screening of Methanolic Extract of Euphorbia hirta linn for Antiinflammatory Activity in Experimental Animals

 

B. Meher1*, T. Satapathy1, A.K .Sahu1, K .K Ahirwar1, P D Kashinath2 and N P Jain3

1Columbia Institute of Pharmacy, Tekari, Raipur C.G. -493111

2Kanakmanjari Institute of Pharmaceutical Sciences, Rourkela -769015

3S.N.D. College of Pharmacy, Yeola, Nasik, India.

Corresponding author: meherbibek@gmail.com

 

 

ABSTRACT:

In the present study an attempt has been made to evaluate the anti inflammatory effect of methanolic extract of Euphorbia hirta Linn in experimentally induced inflammation. The anti inflammatory activity was evaluated using acute inflammatory model like Carrageenan induced paw edema. Oral administration of the extract at the doses 100,200.400mg/kg/b.w exhibited dose dependent and significant anti inflammatory activity in animal models.

 

KEYWORDS: Anti inflammatory, Euphorbia hirta, Carrageenan, Diclofenac.

 


INTRODUCTION:

Nature has provided a complete store-house of remedies to cure all ailments of mankind. In the past, almost all the medicines used were from the plants, the plant being man’s only chemist for ages1. In traditional medicine plants constitute an important source of new biologically active compounds. In India, plants are widely used by all sections of the population either directly as folk medicine or indirectly in the pharmaceutical preparations of modern medicines.  The traditional Indian system of medicine has a very long term history of usage in a number of diseases and disorders but lacks recorded safety and efficacy data2. Inflammation is a local response of living mammalian tissues to the injury it is a body defense reaction in order to eliminate or limit the spread of injurious agents. There are various components to an inflammatory reaction that can contribute to the associated symptoms and tissue injury. Pro inflammatory cytokines (TNF-α and IL-1B) and Nitric oxide (NO) are considered as the key mediators in inflammatory conditions like rheumatoid arthritis, sepsis etc3. As a result of inherent problems associated with the current non steroidal as well as steroidal anti inflammatory agents there is a continuous search especially from natural sources for alternative agents. Large numbers of herbal extract as well as products being employed in the treatment of inflammatory disorders by natural healers4.

 

MATERIALS AND METHODS:

Euphorbia hirta Linn is a annual slender herb belonging to family euphorbiaceae. It is found throughout India in the plains and lower hills, Ceylon, Africa, Australia, and Nigeria etc. 5. The plant material Euphorbia hirta Linn was collected from local region of Yeola and was authenticated by Department of botany. L.K.D.P.P.G. Science College, Dhule, (M.S).

 

Preparation of plant extract:

The shade dried material was coarsely powdered (100gm) and extracted with methanol (1.5 lit.) by cold maceration method. The extract was filtered, concentrated on rotary vacuum evaporator, further dried in vacuum drier and weighed (12.00%w/w). Doses of different concentration of extract (EHM) were dispensed from solution prepared from normal saline. The vehicle alone served as control.

 

Anti Inflammatory Activity:

Drugs and chemicals:

All the drugs and chemicals used in the study were of Pharmaceutical grade. Carrageenan was purchased from Sigma Chemicals Pvt. Ltd, Pune. Diclofenac (Voveran) was purchased from local market used as standard anti-inflammatory drug for comparison study.

 

Animals:

Swiss albino mice of either sex (18-22gm) and Healthy albino rats (Wistar strain) of either sex weighing between 180-220 gm were procured from Yash Farm, Pune. They were maintained under standard conditions of temperature (22±1°C), Relative humidity, 44-56%), 12hr light: dark cycle and fed with standard pellet diet (Amrut, Pranav Agro industries Ltd.) Sangli, India and water ad libitum.  All the animal procedures were performed after approval from the institutional animal ethics committee (IAEC/1167/C/08) and with the permission of Committee for the Purpose of Control and Supervision of Experimental animals (CPCSEA).

 

Acute toxicity studies:

Swiss albino mice of either sex (18-22gm) were used for acute oral toxicity study. The study was carried out as per OECD (Organization of Economic Co-operation and Development) guidelines 423 (Acute toxic class method) and no adverse effects or mortality were observed in the mice up to 5g/kg / p.o. during the 24 hr observation period. Based on the results obtained from the study, the dose for anti-inflammatory activity was fixed to 100, 200, 400 mg/kg for dose dependent study.

 

Grouping of Animals:

The animals (Wister rats) were divided into five groups each containing six animals. Group-I as untreated control and received normal saline , (10ml/kg,p.o) , Group-II served as positive control and received Diclofenac Sodium (5mg/kg/ i.p.) and group –III , IV, and V were treated with 100,200, 400mg /kg EHM  respectively.

 

Method:

Carrageenan induced rat paw edema:6

Effect of extract on Carrageenan induced inflammation was studied in rats according to the methods of winter et.al (1962).  The animals were pre treated with extract (EHM) or Diclofenac Sodium 1hr before the Carrageenan administration.  0.1 ml of 1% Carrageenan solution prepared in normal Saline was injected into sub plantar region of left hind paw of each rat. The paw volume was measured using Plethysmometer before and after 30, 60, 120 and 180 minutes of injection of Carrageenan.

 

Statistical Analysis:

Results were expressed as mean ± S.E.M. The difference between experimental groups was compared by One –way Analysis of Variance (ANOVA) followed by Dunnette’s test. The results were considered statistically significant when P< 0.01.

 

Table -1 Result of acute toxicity study

Sl. no

Dose (mg/kg)

Mortality

Toxic  symptoms

1

300

0/6

None

2

500

0/6

None

3

1000

0/6

None

4

2000

0/6

None

5

5000

0/6

None

 

 

RESULTS:

The methanolic extracts of Euphorbia hirta Linn was evaluated for anti inflammatory activity in acute experimental animal model and the results are summarized in Table-2. The results obtained indicate that the extract showed significant anti inflammatory activity at different test doses i.e. 100,200, 400mg /kg body wt.

 


 

Figure: 1 Effect of different treatments on Carrageenan induced paw edema in rats

 

CONT: Control, DIC: Diclofenac, EHM-Euphorbia hirta methanolic extract.

 

Table -2 Results of anti-inflammatory effect of Euphorbia hirta Linn. against Carrageenan induced paw edema in rats .

Group

Dose (mg/kg)

Initial paw thickness

Paw thickness after 1/2hr

Paw thickness after 1hr

Paw thickness after 2hr

Paw thickness after 3hr

Contrl

10ml/kg

1.188±0.058

1.390±0.062

1.923±0.067

2.020±0.114

2.110±0.085

Diclofenac

5

1.060±0.047

1.230±0.078

1.540±0.083

1.500±0.111**

1.590±0.082*

EHM

100

1.003±0.025

1.208±0.044

1.618±0.046

1.678±0.065

1.628±0.039

EHM

200

1.063±0.043

1.235±0.041

1.573±0.055

1.613±0.055*

1.643±0.044

EHM

400

1.065±0.027

1.233±0.029

1.555±0.058

1.535±0.044**

1.615±0.016

Values are expressed as Mean ±SEM, n=6

Data analyzed by one-way ANOVA followed by Dunnette’s test; **P<0.01,*P<0.05   , EHM ( Euphorbia  hirta   Linn methanolic extract.)

 


DISCUSSION:

In spite  of tremendous development in the field of synthetic drugs during recent era , they are found to have some or other side effects , where as plants still hold their own unique place , by the way of having no side effects . Therefore, a systematic approach should be made to find out the efficacy of plants against inflammation so as to exploit them as herbal anti-inflammatory agents.  It is well known that Carrageenan induced paw edema is characterized by biphasic events with involvement of different inflammatory mediators  .In the first phase (during the first 2h after Carrageenan injection ), chemical mediators such as histamine and serotonin play role , where in second phase (3-4h after Carrageenan  injection ) kinin and prostaglandins are involved 7 . Our results reveled that administration of methanolic extract inhibited the edema at 2 hr in both the dose level i.e. 200 and 400mg .Which is probably inhibition of different aspects and chemical mediators of inflammation.

 

CONCLUSION:

From the above study it is quite apparent that the methanolic extract posses anti-inflammatory activity. Further studies involving the purification of chemical constituents of the plant and the investigation in the biochemical pathway may result in the development potent anti-inflammatory agent with low toxicity and better therapeutic index.

 

REFERENCES:

1.        Kokate  C K,Purohit A P , Gokhale SB, et al. “Pharmacognosy” 45th Edn. Nirali Prakashan Pune, 2010, p.1 .

2.        Jain A, Jain S , Rawat S , Screening of some herbal extracts  for in vitro antifungal activity , Indian Journal of   Nat. Prod., 2010, 26(4), 11 .

3.        Mohan H. “Text book of pathology”, 6th Edn. , Jaypee brother’s medical Publishers Private Limited. New Delhi, 2010,   p. 130 .

4.        Kaneria MS, Naik  S R , Kohli R K ,.Anti-inflammatory ,anti arthritic and analgesic activity of a herbal formulation (DRF/AY/4012)   Indian Journal of Experimental Biology Vol.45, March 2007, p278-84.

5.        Kirtikar  K.R. and Major Basu B.D.   Indian medicinal plants, International book distributors,  Deheradun, 1996,  2, 2197- 98.

6.        Winter CA . Risely E A , Nuss G W , “Carrageenan induced edema in hind paw of the rats as assay for anti-inflammatory drugs”  et al (1962). Proc. Soc. Exptl.Bio.Med. 111:544-47.

7.        Hernandez P M, Rabanal Gallego R M , Anti-inflammatory and analgesic activity of  Sideritis canariensis Var,Pannosa in mice .  (2002).  J. Ethnopharmacol. 81:43-47 .

 

 

 

Received on 09.10.2011          Modified on 20.10.2011

Accepted on 28.10.2011         © RJPT All right reserved

Research J. Pharm. and Tech. 5(1): Jan. 2012; Page 38-40