The present study was designed to carry out the in vitro antioxidant and in vivo nephroprotective activities of Aconitum heterophyllum root ethanolic extract (EEAHR) in acute renal failure induced by glycerol in Wistar albino rats. The antioxidant activity of EEAHR was evaluated by nitric oxide radical scavenging activity (NO assay), hydrogen peroxide (H2O2 assay) and ferric reducing antioxidant power (FRAP) assays. Twenty-four Wistar albino rats were used, divided into four groups of six each to evaluate nephroprotective activity. Group 1 received normal saline, group 2 received hypertonic glycerol (8 ml/kg) alone, and groups 3 and 4 received glycerol + 250 mg/kg EEAHR and glycerol + 500 mg/kg EEAHR, respectively. EEAHR was administered orally 60 minutes before glycerol injection. Kidney damage was measured by serum creatinine, blood urea nitrogen, total protein, albumin, urine output, and histopathological changes. The results showed that the in vitro antioxidant activity was equal to that of vitamin C and in an in vivo study, animals treated with EEAHR showed a significant attenuation of biochemical parameters and histopathological changes in the kidney compared to the glycerol treated group. . more significant with EEAHR 500 mg/kg compared to 250 mg/kg. To conclude, ethanolic extract of Aconitum heterophyllum root showed significant in vitro antioxidant activity and nephroprotective activity in glycerol-induced acute renal failure in Wistar albino rats and is likely to be mediated by the phytochemical constituents and its antioxidant activity. Keywords: nephroprotective, Aconitum heterophyllum root, Antioxidant activity, Glycerol, acute reaction...... middle of paper ......llidine, heteratisin, heterophyllysin, heterophyllidine, 20 α – atisin, 20 β – atisin, atidine, isotisine, ethidine, etsinone and benzoyleteratisine, ajacin [12] and [13]. Ativihsa root mixed honey powder is used for cough irritation and bronchitis. It is also used in non-insulin-dependent diabetes and enteric fevers in infants and children. The leaves of A. heterophyllum have a good aphrodisiac action and a diuretic effect. [14]. Pharmacological actions such as antimicrobial, antifungal, cytotoxic, antioxidant, immunostimulatory and analgesic properties have been reported [15]. It also has anti-inflammatory [16], antidiarrheal [17] and hepatoprotective [18] activities. Therefore, the present study to evaluate the nephroprotective activity of ethanolic extract of Aconitum heterophyllum root in glycerol-induced acute renal failure in Wistar albino rats.