Role of mitochondria and NADPH oxidase derived reactive oxygen species in hyperoxaluria induced nephrolithiasis: therapeutic intervention with combinatorial therapy of N-acetyl cysteine and Apocynin

The interactions between the main cellular sources of ROS, such as mitochondria and NADPH oxidase, are known to play an imperative role in the pathogenesis of hyperoxaluria-induced nephrolithiasis. The present study was designed to investigate the protective effect of a combinatorial therapy based on the attenuation of oxidative stress with antioxidant (N-acetyl cysteine), and NADPH oxidase inhibitor (apocynin), that might be required to effectively eliminate hyperoxaluric manifestations. Hyperoxaluria was induced in male Wistar rats by administering 0.4% ethylene glycol with 1% ammonium chloride in drinking water for 9 days. Hyperoxaluria accentuated renal oxidative stress in terms of increased ROS production and lipid peroxidation. Mitochondria! dysfunction, a central deleterious event in renal stone crystallization, was evident by decreased activities of electron transport chain complex I, II and IV, augmented mitochondrial ROS, reduced GSH/GSSG ratio, which resulted in the mitochondrial permeability transition pore (mPTP) opening as indicated by increased mitochondrial swelling in hyperoxaluric rats. Furthermore, NADPH oxidase activity was significantly increased, with raised expression of NOX1, NOX2, NOX4, p38MAPK and MnSOD, in the renal tissue of hyperoxaluric rats compared to control. However, combinatorial therapy with N-acetyl cysteine (50 mg/kg/day) and apocynin (200 mg/kg/day), intraperitoneally, significantly improved renal functions in hyperoxaluric rats and considerably ameliorated mitochondrial dysfunction. NAC with apocynin was also found to be effective in reducing the redundant activity of NADPH oxidase in renal tissue of hyperoxaluric rats. Hence, our investigation provides novel mechanistic insights that combinatorial approaches using targeted modulators of ROS offer therapeutic benefits in hyperoxaluria-induced nephrolithiasis. (C) 2016 Elsevier B.V. and Mitochondria Research Society. All rights reserved.