- *Corresponding Author:
- R. R. Vohra
Exotic Naturals, 118 Mourya House, Link Road, Andheri (W), Mumbai-400 053, India
E-mail: [email protected]
|Date of Submission||14 April 2008|
|Date of Revision||30 April 2008|
|Date of Acceptance||4 December 2008|
|Indian J. Pharm. Sci., 2008, 70 (6): 785-787|
The antiinflammatory activity of the polyherbal formulation Entox® was investigated in rats for acute and sub acute models of inflammation using carrageenan-induced rat paw edema and cotton pellet granuloma methods respectively at a dose of 300 mg/kg and 600 mg/kg administered orally. The formulation in doses of 300 mg/kg and 600 mg/kg showed 51.61% and 54.84% inhibition of paw edema, respectively at the end of 3 h. The percent inhibition of granuloma by cotton pellet method was 27.92% and 53.17%, respectively. The formulation showed a significant antiinflammatory activity in both the experimental models and the activity was comparable to that of the standard drug, indomethacin.
Entox®, carrageenan, cotton pellet granuloma, antiinfl ammatory, polyherbal, formulation
The formulation, Entox® is constituted of fruits of Terminalia chebula (Combretaceae) and Embelica officinalis (Euphorbiaceae), fruit rind of Punica granatum (Punicaceae), bark of Terminalia arjuna (Combretaceae), roots of Rubia cordifolia (Rubiaceae), Withania somnifera (Solanaceae) and Tinospora cordifolia (Menispermaceae) and rhizomes of Curcuma longa (Zingiberaceae). Entox® is currently being used as an antioxidant formulation. The studies have shown that oxidation is one of the mechanisms in the inflammatory process . The antiinflam onstituents of this formulation which includes Terminalia chebula , Emblica officinalis , Punica granatum , Terminalia arjuna , Rubia cordifolia , Withania somnifera , Tinospora cordifolia  and Curcuma longa  has been reported in scientific literature. Hence the present study was designed to study the antiinflammatory activity of all the constituents when combined together.
The antiinflammatory activity was evaluated in male albino rats of Wistar strain (150-200 g) by both carrageenan-induced rat paw edema and cotton pellet granuloma method. All the experimental protocols were approved by Institutional Animal Ethics Committee (IAEC). The rats were given a standard laboratory diet and water ad libitum. In carrageenan-induced rat paw edema method, the animals were divided into 4 groups of 6 animals each. Food was withdrawn 16 h before drug treatment. The first group received 0.5 ml of 1% gum acacia solution orally and served as a control. The second group of animals was administered indomethacin (10 mg/kg, p.o.). The animals of groups 3 and 4 were treated with the formulation (300 mg/kg and 600 mg/kg, orally). Acute inflammation was produced by sub plantar injection of 0.1 ml of 1% suspension of carrageenan in normal saline, in the left hind paw of the rats, 1 h after oral drug treatment. The paw volume was measured plethysmometrically at 0, 1, 2, 3 and 4 h after the carrageenan injection. The difference in initial and the subsequent reading gave the actual edema volume and the percent inhibition was calculated [10,11] (Table 1). The data was analyzed using one way ANOVA followed by Dunnetts test and p<0.01 was considered significant. In cotton pellet granuloma method, sterile cotton pellets (30±1 mg) were implanted subcutaneously bilaterally in the groin region under light ether anesthesia. The animals were divided into 4 groups of 6 animals each and were treated consecutively for 7 d. The treatment for all four groups was the same as that mentioned in the earlier method except that indomethacin (5 mg/ kg, orally) was administered to second group. The animals were sacrificed on the 8th d. The granulation tissue with cotton pellets was removed and dried overnight at 60û to constant weight. The difference between dry weight of dissected cotton pellet and the weight of individual cotton pellet before implantation gave the dry weight of granuloma [11,12] (Table 2). The data was analyzed using one way ANOVA followed by Dunnetts test and p<0.01 was considered significant.
|Time (h)||Volume of edema (ml)|
|Control (Vehicle, p.o. )||Indomethacin (10 mg/kg, p.o. )||Formulation (300 mg/kg, p.o. )||Formulation (600 mg/kg, p.o. )|
|1||0.55±0.03||0.33±0.06* (40.00)||0.38±0.03 (31.82)||0.30±0.05* (45.56)|
|2||0.65±0.04||0.35±0.05* (44.00)||0.43±0.02* (32.00)||0.38±0.03* (40.00)|
|3||0.78±0.06||0.30±0.05* (61.29)||0.38±0.03* (51.61)||0.35±0.05* (54.84)|
|4||0.68±0.06||0.28±0.04* (59.26)||0.38±0.03* (44.44)||0.33±0.02* (51.85)|
Values are expressed as mean±SEM, n=6 in each group. Values in parentheses indicate percent inhibition of paw edema.*P< 0.01 as compared with control.
Table 1: Effect of formulation on carrageenan –induced rat paw edema
|Treatment||Dry weight of granuloma† (mg)|
|Control (Vehicle, p.o.)||48.80±0.89 (-)|
|Indomethacin (5 mg/kg, p.o.)||20.88±0.63* (57.08)|
|Formulation (300 mg/kg, p.o.)||35.13±0.51* (27.92)|
|Formulation (600 mg/kg, p.o.)||22.78±0.43* (53.17)|
Difference between weight of dissected dry cotton pellet and weight of cotton pellet before implantation. Values are expressed as mean±SEM, n=6 in each group. Values in parentheses indicate percent inhibition of granuloma formation.*P< 0.01 as compared with control.
Table 2: Effect of formulation on cotton pellet granuloma in rats
The antiinflammatory effect of the formulation on carrageenan-induced rat paw edema is presented in Table 1. The development of carrageenan-induced edema is biphasic, the 1st phase is mediated through the release of histamine and serotonin, with a peak value at 1h; whereas the 2nd phase is related to the release of prostaglandins with a peak value at 3 h [13,14]. Edema suppressant effect of 300 mg/ kg and 600 mg/kg treated groups were found to be significant on both the phases of inflammation when compared to control. The 300 mg/kg and 600 mg/kg doses showed maximum anti-edematous effect of about 51.61% and 54.84%, respectively at 3 h after carrageenan administration which was found to be significant statistically (P<0.01) as compared to control. Indomethacin (standard) showed similar type of reduction at 3 h (P<0.01) as compared to control. Since the formulation in our study exhibited significant inhibitory effect on the rat paw edema, it probably exerts an inhibitory effect on some of the mediators of inflammation induced by the carrageenan stimuli. Swingle and Shildeman demonstrated that there are three phases of inflammation after pellet implantation ; consisting of a transudative phase, defined as the increase in wet weight of the pellet which occurs during the first 3 h (passive process), an exudative phase which occurs between 3 and 72 h after implanting the pellet (a delayed-prolonged vascular permeability change in inflammation) and a proliferative phase, measured as the increase in dry weight of granuloma which occurs between 3 and 6 d after implantation . In our study, the antiinflammatory activity was indicated by a significant reduction in the granular tissue formation. The animals when treated with formulation at doses of 300 mg/kg and 600 mg/kg orally showed a significant (P<0.01) inhibition of granuloma formation of 27.92% and 53.17%, respectively and indomethacin (5 mg/kg, p.o.) showed 57.08% inhibition as compared to control. In conclusion the formulation exhibits remarkable antiinflammatory activity in the acute phase as well as in the subacute model of inflammation.
- Kapoor M, Clarkson AN, Sutherland BA, Appleton I. The role of antioxidants in models of inflammation: Emphasis on l-arginine and arachidonic acid metabolism. Inflammopharmacol 2005;12:505-19.
- Rasool M, Sabina EP. Antiinflammatory effect of the Indian Ayurvedic herbal formulation Triphala on adjuvant-induced arthritis in mice.Phytother Res 2007;21:889-94.
- Asmawi MZ, Kankaanranta H, Moilanen E, Vapaatalo H. Antiinflammatory activities of EmblicaofficinalisGaertn leaf extracts. J Pharm Pharmacol 1993;45:581-4.
- Al-Yahya MA. Preliminary Phytochemical and Pharmacological Studies on the Rind of Pomegranate (Punicagranatum L.). Pak J Bio Sci 2005;8:479-81.
- Tripathi YB, Reddy MM, Pandey RS, Subhashini J, Tiwari OP, Singh BK, et al. Antiinflammatory properties of BHUx: A polyherbal formulation to prevent atherosclerosis. Inflammopharmacol 2004;12:131- 52.
- Kasture SB, Kasture VS, Chopde CT. Antiinflammatory activity of Rubiacordifolia roots. J Nat Rem 2001;1:111-5.
- Rasool M, Varalakshmi P. Immunomodulatory role of Withaniasomnifera root powder on experimental induced inflammation: An in vivo and in vitro study. VasculPharmacol 2006;44:406-10.
- Utpalendu J, Rabindranath C, Badriprasad S. Preliminary studies on antiinflammatory activity of Zingiberofficinalerosc.,Vitexnegundolinn. andTinosporacordifolia (willid) miers in albino rats. Indian J Pharmacol 1999;31:232-33.
- Kohli K, Ali J, Ansari MJ, Raheman Z. Curcumin: A natural antiinflammatory agent. Indian J of Pharmacol 2005;37:141-7.
- Winter CA, Risley EA, Nuss GW. Carrageenan induced oedema in hind paw of the rats as an assay for antiinflammatory drugs. ProcSocExpBiol 1962;111:544-7.
- Weithmann KU, Bartlett RR, Schieyerbach R. Analgesic, antiinflammatory and antipyretic activity. In: Vogel GH, editors. Drug discovery and Evaluation, Pharmacologicl Assays 2nd ed. Hiedelberg: Springer; 2001. p. 767-9.
- Winter CA, Risey EA, Nuss GW. Antiinflammatory and antipyretic activities of indomethacin, 1-(p-chlorobenzoyl)-5-methoxy-2- methylindole-3-acetic acid. J PharmacolExpTher 1963;141:369-76.
- Vinegar R, Schreiber W. Biphasic development of carrageenan edema in rats. J PharmacolExpTher 1969;166:95-103.
- Bhatt KG, Sanyal RK. Association of histamine and 5-hydroxytryptamine with the inflammatory processes. J Pharm Pharmacol 1962;15:78-9. 15.
- Swingle KF, Shildeman FE.Phase of inflammatory response to subcutaneous implantation of a cotton pellet and their modification by certain antiinflammatory agents. J PharmacolExpTher 1972;183:226- 34.