The anti-inflammatory effect of a petroleum ether extract of Vitex negundo Linn (Verbenaceae) leaves, (PEVNL) (250 and 500 mg/kg, p.o.), was evaluated in carrageenan-induced hind paw edema and cotton pellet granuloma models. Diclofenac sodium was used as a standard drug. The biochemical parameters were estimated in serum. In carrageenan-induced rat paw edema model PEVNL exhibited significant (p < 0.01) inhibition of edema volume at 4 h in a dose-dependent manner. In the cotton pellet granuloma model, dry weight of cotton pellets was significantly inhibited in a dose-dependent manner with maximum effect noticed at 500 mg/kg, p.o. Both doses of PEVNL were found to normalize the increased alkaline phosphatase, acid phosphatase, alanine amino transferase (ALT) and aspartate amino transferase (AST) and reversed the decreased serum albumin. In conclusion, PEVNL exhibited anti-inflammatory activity in acute and subacute models. At a dose of 500 mg/kg, p.o., PEVNL was found to possess maximum activity, and this effect was comparable with reference drug diclofenac sodium (5 mg/kg, p.o.).