Elsevier

Toxicon

Volume 36, Issue 1, 1 January 1998, Pages 207-215
Toxicon

Viper venom-induced inflammation and inhibition of free radical formation by pure compound (2-hydroxy-4-methoxy benzoic acid) isolated and purified from anantamul (Hemidesmus indicus R.Br) root extract

https://doi.org/10.1016/S0041-0101(97)00070-6Get rights and content

Abstract

The present investigation explored the possible venom neutralizing effect of a pure compound (2-hydroxy-4-methoxy benzoic acid) isolated and purified from the methanolic root extract of Hemidesmus indicus R.Rr. 2-OH-4-MeO benzoic acid possessed potent anti-inflammatory, antipyretic and antioxidant properties. The compound effectively neutralized inflammation induced by Vipera russelli venom in male albino mice and reduced cotton pellet-induced granuloma in rats. The compound produced a significant fall in body temperature in yeast-induced pyrexia in rats but did not change the normothermic body temperature. The compound effectively neutralized viper venom-induced changes in serum phosphatase and transaminase activity in male albino rats. It also neutralized free radical formation as estimated by TBAPS and superoxide dismutase activities. The antisnake venom activity of the pure compound is partly mediated through the above physiological process.

Introduction

In India, many plants are reported to be used as a remedy for a number of diseases. The root of Hemidesmus indicus R.Br, locally called anantamul, is found throughout India. It is used for the treatment of many diseases as a blood purifier, diuretic, antirheumatic and antidote in snakebite (Chopra et al., 1956; Alam et al., 1996). Despite the traditional use of the plant in folk medicine, only a few articles have reported the chemical constituents of the H. indicus species, including coumarino-lignoids, flavonoids and triterpenoids (Mandal et al., 1991), and knowledge of the pharmacological action of its extracts and constituents remains poor. Studies from this laboratory have recently demonstrated an active antisnake venom compound isolated, purified and characterized from the methanolic root extract of H. indicus (Alam et al., 1994). The present investigation explored the snake (Russell's viper) venom neutralizing activity of the pure compound (2-hydroxy-4-methoxy benzoic acid) on experimental rodents.

Section snippets

Venom

Lyophilized snake venom from Russell's viper (Vipera russelli) was commercially collected from Calcutta Snake Park (Calcutta, India) and preserved in a desiccator at 4°C in an amber-coloured glass vial until further use.

The snake venom was dissolved in 0.9% saline and centrifuged at 900 g for 10 min. The supernatant was used as venom and kept at 8°C until further use. The venom concentration was expressed in terms of dry weight (mg/ml, stock venom solution).

Animals

Charles Foster albino rats (male),

Anti-inflammatory activity of pure compound

The anti-inflammatory activity of the pure compound was assessed by mouse paw oedema and cotton pellet granuloma. Mouse paw oedema induced by V. russelli venom (1 μg) attained its peak at 1 hr of observation. The pure compound at a dose of 5 mg/kg (i.p.) was found to produce significant inhibition of venom-induced inflammation. Inhibition of inflammation induced by the pure compound was maximum (53.7±1.7%) at 2 hr of observation, as compared with aspirin (44.7±0.95%) and indomethacin (40.2±1.7%).

Discussion

Injury and death due to snakebite are sociomedical problem that occur throughout the world. Snake envenomation causes different pathophysiological changes such as inflammation, increased body temperature, haemorrhage, necrosis, nephrotoxicity, cardiotoxicity, haemostatic changes and ultimately death (Theakston and Reid, 1983). Antiserum is the only therapeutic agent that will neutralize venom, despite several side-effects. The search for suitable antagonists against snake envenomation is

Acknowledgements

The authors are grateful to the Council of Scientific and Industrial Research, New Delhi, India, for partial financial support (Senior Research fellowship to MIA).

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