The present study aimed to achieve the in vitro antioxidant and in vivo nephroprotective activities of the ethanolic extract of Aconitum heterophyllum root (EEAHR) in the glycerol-induced acute renal failure in Wistar. albino rats. The antioxidant activity of EEAHR was evaluated by 2,2-diphenyl-picrylhydrazyl scavenging activity (DPPH assay), nitric oxide radical scavenging (NO assay), hydrogen peroxide assays. (H2O2 dosage) and ferric reducing antioxidant power (FRAP). Twenty-four Wistar albino rats were used and 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 of EEAHR respectively. EEAHR was administered orally 60 min before the glycerol injection. Renal injury 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, EEAHR-treated animals showed significant attenuation of biochemical parameters and histopathological changes of the kidney compared to the glycerol-treated group. more significant with EEAHR500 mg/kg than with 250 mg/kg. To conclude, the ethanolic extract of Aconitum heterophyllum root showed significant in vitro antioxidant activity and nephroprotective activity in glycerol-induced acute renal failure in albino Wistar rats and it is likely to be mediated by phytochemical constituents and its antioxidant activity. Keywords: Nephroprotector, Aconitum heterophyllum root, Antioxidant activity, Glycerol, re...... middle of paper ......llidine, heteratisine, heterophyllisine, heterophyllidin, 20 α – atisine, 20 β – atisine, atidine, isotisine, hetidine, hetsinone and benzoylheteratisine, ajacin [12] & [13]. Honey mixed with ativihsa root powder is used for cough irritations and bronchitis. It is also used in non-insulin-dependent diabetes and enteric fevers in infants and children. A. Heterophyllum leaves have a good aphrodisiac action and a diuretic effect. [14]. The pharmacological actions reported are antimicrobial, antifungal, cytotoxic, antioxidant, immunostimulating and analgesic properties [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 albino Wistar. rats.