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Anti-nociceptive and Anti-inflammatory Activities of Wrightia Arborea

Laizuman Nahar
Department of Pharmacy, University of Rajshaho, Rajshahi - 6025, Bangladesh

Fatema Nasrin
Department of Pharmacy, Southeast University, Dhaka - 1213, Bangladesh

Ronok Zahan
Department of Pharmacy, University of Rajshaho, Rajshahi - 6025, Bangladesh

Md. Ashik Mosaddik
Department of Pharmacy, University of Rajshaho, Rajshahi - 6025, Bangladesh

Abstract/ Executive Summary:

Anti-nociceptive and anti-inflammatory effects of methanolic extract of Wrightia arborea (MEWA) were examined using different models in rats. MEWA was given to rats orally upto 2000 mg kg^-1 b.wt. for acute toxicity study and observed for 14 days. Anti-nociceptive activity was evaluated in rats against Acetic acid induced writhing (chemically induced pain) and Tail immersion method (thermally induced pain). Acute anti-inflammatory activity of MEWA was also evaluated in Formaline-induced rat paw edema model and Carrageenan-induced hind paw edema model in rats. Results demonstrated that no mortality was found upto single dose of 2000 mg kg^-1 b.wt. in rats even after 14 days observation. In comparison to control group MEWA at 100 and 200 mg kg^-1 b.wt. showed highly significant anti-nociceptive activity against chemically (p<0.001) as well as thermally (p<0.05 and p<0.001) induced pain as compared to standard drugs, indomethacin and nalbufin, respectively. In the formalin test, both the doses of 100 and 200 mg kg^-1 of extract significantly prevented increase in volume of paw edema (p<0.05 and p<0.01) both in the neurogenic and inflammatory phases. MEWA (200 and 400 mg kg^-1 p.o.) also significantly prevented increase in volume of paw edema in Carrageenan test (p<0.05 and p<0.001). The results suggest that MEWA has significant analgesic and anti-inflammatory potential which may be mediated by central and peripheral mechanism.

Keywords: Anti-nociceptive, Anti-inflammatory, Writhing, Tail immersion test, Carageenan

Location:

Start Date: 00-00-2011

End Date: 00-00-2011

Program Area: Development of Host and Medicinal Plants

Commodity: Plant(s)

Objectives:

To assess potential of the methanolic extract of W. arborea leaves as anti-nociceptive and anti-inflammatory agents.

Methodology:

Collection and Identification of plant: The leaves of Wrightia arborea were collected from Hill tracts, Chittagong, Bangladesh in July 2011. The Voucher Specimen No.147 is kept for W. arborea in Herbarium of the Department of Botany, University of Chittagong, Bangladesh. Preparation of extract: The collected aerial parts of plant was cleared, dried under shade at room temperature and powdered. For the extraction of phytochemicals about 200 g powder of the plant was taken in a glass jar and completely submerged with 600 mL methanol for 7 days accompanying occasional shaking and stirring. The extract was then filtered through filter paper (Double Rings filter paper 102, 11.0 cm). The filtrates was concentrated at 50°C under reduce pressure using vacuum pump rotary evaporator (STUART RF3022C, UK) to yield a solid residue 30 g and stored at 4°C in air tight containers for further studies. Design of experiment - Experimental animal: Experiment was conducted on adult albino rats of male sex with the weights of 120-140 g procured from International Centre for Diarrheal Disease Research Bangladesh (ICDDRB). All rats were fed normal laboratory chow food containing 16% protein, 66% carbohydrate, 8% fats and water. All rats were housed at a (12:12) h light and dark cycle at 25°C and relative humidity (60-70) %. Ethical approval: The guidelines followed for animal experiment were accepted by the institutional animal ethical committee. Drugs and chemicals: Indomethacin and Nalbufin were the gift samples from Square pharmaceuticals ltd, Bangladesh. Formalin and Acetic acid were obtained from BASF, Bangladesh. And all other reagents used were of analytical grade. Acute toxicity test: Acute toxicity study was performed for the extract according to the acute toxic classic method. Extract was given orally to rats at the graded doses like 100, 300, 1000 and 2000 mg kg^-1 b.wt. Immediately, after dosing, the animals were observed continuously for first four hours for behavioral change and mortality for daily upto 14 days. Analgesic activity - Acetic acid induced writhing method: The peripheral anti-nociceptive activity of MEWA was studied using acetic acid-induced writhing model in rats. The abdominal writhing was induced by intraperitoneal injection of acetic acid solution (0.7%, 0.1 mL 10^-1 g b.wt.) to each rat, a model of visceral pain. Five groups of mice (n = 6) were randomly formed. Indomethacin (10 mg kg^-1) was used as standard analgesic agent. MEWA was administered at 100 and 200 mg kg^-1 b.wt. were orally administered 30 min prior to the injection of acetic acid. The number of writhing was calculated for 10 min after the application of acetic acid. The percent inhibition (% analgesic activity) was calculated by:

Inhibition (%) = {(A-B)/A}×100

where, A is average number of writhing of the control group, B is average number of writhing of the test group.

Tail immersion method: This study is a thermal method to assess analgesia according to the method of Luiz. Rats divided in the groups of six each, were held in position in a suitable restrainer with the tail extending out. About 3-4 cm area of the tail was marked and immersed in the water bath thermo-statistically maintained at 51°C. The withdrawal time of the tail from hot water (in seconds) was noted as the reaction time or tail flick latency. 0.2 mL of 0.9% NaCl solution was administered to control animals, MEWA in doses of 100 and 200 mg kg^-1 were given orally by intubation. The initial reading was taken immediately before administration of test and standard drugs (Nalbufin, 10 mg kg^-1) and then 30, 60 and 90 min after the administration. The criterion for analgesia was postdrug latency which was greater than two times the pre-drug average latency. Antinflammatory activity: Formalin-induced paw edema the antinflammatory activity of MEWA was determined using the formalin tes. Control group received 5% formalin. Twenty μL of 5% formalin was injected into the dorsal surface of the right hind paw 60 min after administration of MEWA (100 and 200 mg kg^-1, b.wt.) and indomethacin (10 mg kg^-1, b.wt.). The increase in paw diameter was measured using vernier caliper. The difference in edema of the right hind paw and the left hind paw indicates inflammation. Measurement was done immediately before and after 1-5 h following formalin injection. Carrageenan induced paw edema: The animals were divided into five groups. Acute inflammation was produced by subplantar injection of 0.1 mL of 1% suspension of carrageenan with 2% gum acacia in normal saline, in the right hind paw of the rats, 1 h after oral administration of MEWA at dose of 200 and 400 mg kg^-1, b.wt. The paw volume was measured at 1, 2, 3 and 4 h after the carrageenan injection using micrometer screw gauge. The difference between the two readings was taken as the volume of edema. Indomethacine suspended in 2% gum acacia was used as the standard drug. Statistical analysis: Data were analyzed by one-way ANOVA followed by Dunnett’s test. Value p<0.05 was considered statistically significant.

Source of Information: Pakistan Journal of Biological Sciences, 16: 485-490; ISSN 1028-8880

Article Link (Online Journal): DOI: 10.3923/pjbs.2013.485.490; URL: http://scialert.net/abstract/?doi=pjbs.2013.485.490

Funding Source:

Budget:

Total Budget (Tk.) :

Achievement/Findings:

On the basis of results obtained from the present study, it can be concluded that the plant extract possesses remarkable analgesic and anti-inflammatory activities. Present work was a preliminary effort which will require further detailed investigation including characterization of active compounds and requires preformulation studies for development of a potential dosage form.

Knowledge Management: Journal