International Journal of Engineering Technology Science and Research IJETSR www.ijetsr.com ISSN 2394 – 3386 Volume 2 Issue 2 February 2015
Anti-Inflammatory Activity of the Stem and the Root Extracts of Alangium Salviifolium Decapetalum
T. Raga Sudha
Dept. of Microbiology, Indirapriyadarshini Govt. Degree College, Nampally, Hyderabad 500 001 T.S. India.
Abstract: Methanolic (95%) extracts of the stem and roots of Alangium salviifolium Decapetalum were screened for anti-inflammatory activity in carrageenan induced rat paw oedema. The results in the experimental were found to be significant when compared to control. On comparison, the root and stem extracts had exhibited anti-inflammatory effect at the same potency.
Key words: Alangium salviifolium, antiinflammatory activity, stem, root, methanol extract, carrageenan-induced paw oedema method.
INTRODUCTION The family Alangiaceae consists of 22 species out of which Alangium salviifolium Decapetalum is mainly used as medicine in India, China, Burma, and Philippines (Hooker J.D., 1875). Alangium salviifolium is a deciduous tropical tree with thorny leaves. The different parts of this plant were reported to possess acrid, astringent, emollient, anti-helmintic, diuretic and purgative properties (Kirtikar & Basu, 1994). It is also used externally in acute cases of rheumatism, inflammation and leprosy. It is used externally and internally in case of rabid dog bite. The root bark is an antidote for several poisons. Fruits are used as a poultice for treating burning sensation and hemorrhage (Kirtikar & Basu, 1994). The leaves are poultice to reduce rheumatic pains. The stem bark is used both as contraceptive and abortifacient (Murugan, et.al., 2000) and also used in vomiting and diarrhea. In this study the root (ASRE) and stem (ASSE) extracts along with their barks were studied for their anti-inflammatory activity, as they were known to be in use traditionally.
MATERIAL AND METHODS Plant material The plant material for the study was collected from the outskirts of Visakhapatnam district, Andhra Pradesh in the month of November 2005 and was authenticated by Prof. M. Venkaiah, Dept of Botany, Andhra University, Visakhapatnam, Andhra Pradesh, INDIA. A voucher specimen (BGR- AS1) was deposited in the herbarium of Andhra University, Visakhapatnam.
Extraction The stems (500g) and roots (500g) of Alangium saliviifolium were shade dried, pulverized into coarse powder and extracted in Soxhlet apparatus with methanol 95%. The extract was concentrated under reduced pressure, dried completely and weighed (Anonymous Pharmacopoeia of India, 1980). The percentage yield of methanolic ASRE and ASSE are 6gm and 8gm respectively. The extract was stored in a refrigerator and a weighed quantity was suspended in 1% sodium carboxy methyl cellulose (CMC) solution for the experiment.
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International Journal of Engineering Technology Science and Research IJETSR www.ijetsr.com ISSN 2394 – 3386 Volume 2 Issue 2 February 2015
Preliminary Biochemical test The phytochemical tests of the stem extract showed the presence of steroids, terpenoids, alkaloids, carbohydrates, flavonoids, and tannins. The phytochemical investigation of the root extract showed the presence of steroids, alkaloids and terpenoids.
Animals Used Colony bred adult albino Wister rats of either sex (supplied by B.N. Ghosh & Co., Calcutta) weighing 200g – 250g were selected for the study. Animals were maintained at 250C ± 20C, relative humidity 50 ± 15% under 12hr light/dark cycle. The animals were fed with standard pellet diet (Ratan Brothers, India) and water ad libitium. The experimental protocol has been approved by the Institutional Animal Regulatory body of the Government (Regd. No. 516/01/A/CPCSEA).
Anti-Inflammatory activity The anti-inflammatory activity of the methanolic stem and root extracts of Alangium saliviifolium was determined in Wister strain albino rats using carrageenan-induced paw oedema method (Winter et. al.,1962). The rats were divided into eight groups (I, II, III, IV, V, VI, VII, VIII), four in each group. Group I was treated with drug vehicle, 1% sodium CMC in saline (-ve control); Group II was treated with non-steroidal anti-inflammatory drug ibuprofen (+ve control); Group III, IV, V received crude methanolic ASSE (200, 400, 800mg/kg body weight) while the rest of the groups received crude ASRE ( 200, 400, 800mg/kg body weight). All doses (0.1ml/100g body weight) were administrated orally 1h prior to the induction of oedema. Then 0.1ml of 1% carrageenan solution was injected into the subplantar surface of the right hind paw of the rats. Simultaneously the left paws were injected with saline water. The thickness of the rats paw was measured using Zeitlin’s Constant loaded Lever apparatus (Battu et.al., 2000). The paw thickness of the rats was measured for 6h with 1h interval. Zero hour readings are the initial paw reading of the animals. The degree of paw oedema was calculated as follows: % increase in paw thickness = ((Y1-Y0)/ Y0) x 100 Where Y1 = paw thickness at time t (1, 2, 3, 4, 5, 6hr) after injection Y0 = paw thickness at 0 hr i.e. before injection Therefore % increase due to carrageenan = (% increase in right paw - % increase in left paw).
RESULTS The edema and percentage of rate of inhibition for each group were calculated. From the observed values, the percentage of maximal paw oedema produced during 6h was calculated and plotted as line graph. The percentage of total paw oedema as area under the time course curve (AUC) produced during 6h was also calculated and plotted as bar graph. The data obtained from the experiment were expressed as means ± S.E.M, N = 4. Student’s t-test was used to compare test and control group values. The differences in the test verses control values were considered statistically significant at P ≤ 0.05. The Alangium saliviifolium root and stem extracts produced dose related significance (P < 0.05 – 0.001). DISCUSSION In comparison with the drug vehicle-control group, all the test groups had shown a significant reduction in inflammation. The root and the stem extracts has shown the same potency. The percentage of inhibition of edema by 800mg dosage of stem (89.001.0) and root (89.000.5) extract was almost equal to that of the standard drug, ibuprofen (89.021.0). According to Vineger et. al., 97 T. Raga Sudha
International Journal of Engineering Technology Science and Research IJETSR www.ijetsr.com ISSN 2394 – 3386 Volume 2 Issue 2 February 2015
(1987) the development of the carrageenan induced edema derives from the release of cytoplasmic enzymes and inflammatory mediators like histamine and serotonin from mast cells and the increase of prostaglandin and thromboxanes via the catalytic activity of cycloxygenase enzymes (COX-1 & COX-2) in the inflammatory area (Vineger et. al., 1987). The macrophages in the carrageenan induced dermal tissue releases much interlukin-1 to induce accumulation of polymorphic nuclear cells (PMN) into the inflammatory area. The activated PMNs then release the lysozomal enzymes and activate oxygen, especially superoxide dismutase, to destroy connective tissue and induce paw swelling. The suppression of increase in paw oedema by ibuprofen is due to the inhibition of the enzyme arachidonate cyclooxygenase or COX (Rang et. al.,1999). The results of present study suggest that the methanol extracts of Alangium salviifolium at a dose of 200, 400 and 800 mg/kg significantly (p<0.001) suppressed carrageenan-induced rat paw oedema. The observed anti-inflammatory effect of Alangium saliviifolium extracts could be due to the presence of biologically active chemical compounds like steroids, flavonoids, terpenoids and alkaloids in the extract, which are known to possess anti-inflammatory activity (Jackson & Jason, 2001). Further studies are necessary to elucidate the mechanism and phytochemical responsibly for anti-inflammatory activity. Acknowledgement I express my deep sense of gratitude to Prof. B. Ganga Rao, Principal, A.U. College of Pharmacieutical Sciences, Andhra University, Visakhapatnam, Andhra Pradesh, for his encouragement and guidance in executing the experimental.
Table 1 : Percentage inhibition of carrageenan-induced paw oedema in rats by prophylactic treatment with methanolic extract of Alangium salviifolium stem and Ibuprofen.
S.No. Treatment Percentage inhibition of Percentage inhibition of the maximal paw the total (AUC) paw oedema during 6h oedema during 6h 1 Group A (Drug Vehicle) 0.05.2 0.00.9 2 Group B (Ibuprofen) 81.61.8 89.021.0 3 Group C (ASS200) 56.9354.0 59.790.7 4 Group D (ASS400) 67.8251.8 71.380.4 5 Group E (ASS800) 86.680.1 89.001.0
Table 2 : Percentage inhibition of carrageenan-induced paw oedema in rats by prophylactic treatment with methanolic extract of Alangium salviifolium root and Ibuprofen.
S.no. Treatment Percentage inhibition of Percentage inhibition of the maximal paw the total (AUC) paw oedema during 6h oedema during 6h 1 Group A (Drug Vehicle) 0.04.3 0.073 2 Group B (Ibuprofen) 77.9718 89.021.2 3 Group C (ASR200) 40.863.6 59.794.2 4 Group D (ASR400) 67.742.7 71.380.4 5 Group E (ASR800) 86.840.9 89.000.5 98 T. Raga Sudha
International Journal of Engineering Technology Science and Research IJETSR www.ijetsr.com ISSN 2394 – 3386 Volume 2 Issue 2 February 2015
Graph 1
Mean s.e.m., N=4
70 Control (Sodium CMC) 60 Ibuprofen5mg 50 ASR 200mg 40 ASR400mg ASR800mg 30 20 10 0
% increase %in increase rat oedema paw 0 1 2 3 4 5 6 7 Hour(h)
Effects of thee crude extract of Alangium salviifolium root(ASR) (200, 400, 800mg.kg-1) and the standard drug Ibuprofen(2.5 10-5mol.kg-1 body weight) on the maximal paw oedema in carrageenan-induced rats. Significance: significance=***p<0.001,ns=not significant
Graph 2
110 Mean s.e.m, N=4 100 90 Drug Vehicle 80 Ibuprofen 5mg 70 ASR200mg 60 ** ASR400mg 50 ASR800mg 40 30 *** 20 *** 10 ***
mean mean control response 0 Total oedema (AUC)Total oedema % as of
Effect fo the crude extract of Alangium salviifolium root(ASR) (200, 400, 800mg.kg-1) and the standard drug Ibuprofen(2.510-5mol.kg-1body weight) on the total paw oedema in carrageenan-induced rats. Significance: significance=***p<0.001;ns=not significance
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International Journal of Engineering Technology Science and Research IJETSR www.ijetsr.com ISSN 2394 – 3386 Volume 2 Issue 2 February 2015
Graph 3
Mean s.e.m., N=4
70 Drug Vehicle 60 Ibufropen5mg 50 ASS 200mg 40 ASS400mg ASS 800mg 30 20 10 0
% increase %in increase rat oedema paw 0 1 2 3 4 5 6 7 Hour(h) Effect of the crude extract of Alangium salviifolium stem(ASS) (200, 400,800mg.kg-1) and the standard drug Ibuprofen (2.5 X 10-5mol.kg-1body weight) on the maximal paw odma in carrageenan-inducd paw oedema. Significance: significance= ***p<0.001, ns = not significance
Graph 4
Mean s.e.m, N=4
110 100 Drug vehicle 90 Ibuprofen5mg 80 ASS200mg 70 60 ASS400mg 50 *** ASS800mg 40 *** 30 20 *** *** 10
mean mean control response 0 Total oedema Total(AUC) oedema %as of
Effects of the crude extract of Alangium salviifolium stem(ASS) (200, 400, 800mg.kg-1) and the standard drug Ibuprofen(2.5 10-5mol.kg-1 body weight) on the total paw oedema in carrageenan-induced rats. Significance: significance=***p<0.001,ns=not significant
100 T. Raga Sudha
International Journal of Engineering Technology Science and Research IJETSR www.ijetsr.com ISSN 2394 – 3386 Volume 2 Issue 2 February 2015
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