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Background: Hyperoxaluria and calcium oxalate crystal deposition in the kidneys lead to overproduction of reactive oxygen species, resulting in the development of oxidative stress and renal injury. At the cellular levels, mitochondria and NADPH oxidase involved in reactive oxygen species production play a crucial role in the pathogenesis of hyperoxaluria-induced renal injury. Objective: The objective was to investigate the therapeutical effect of homoeopathic drug Berberis vulgaris, a potent antioxidant, upon regulation of NADPH oxidase against acute high-grade sodium oxalate-induced hyperoxaluria in rats. Materials and Methods: Hyperoxaluria was induced in male Wistar rats by administering a single dose of sodium oxalate (70 mg/kg body weight) intraperitoneally, and the treatment groups were pre-treated with homoeopathic drug Berberis vulgaris 6c (20 μl/100 g of body weight) ultra low dose for 7 days. Results: Berberis vulgaris significantly reduces hyperoxaluria-induced oxidative stress and restores antioxidant enzyme activities in kidney tissue. Histological analysis depicted that Berberis vulgaris treatment decreases renal epithelial damage and inflammation and restored normal glomerular morphology. Furthermore, immunoblotting analyses of NADPH oxidase revealed significant increased activity in the renal tissue of hyperoxaluric rats when compared to that of control rats. This has been brought back to normal by Berberis vulgaris treatment. Conclusion: Thus, our results emphasised that the Homoeopathy drug Berberis vulgaris has been effective in ameliorating sodium oxalate-induced acute hyperoxaluria in Wistar rats by modulating mitochondrial oxidative stress through the inhibition of NADPH oxidase.