Evidence against increased hydroxyl radical production during oxygen deprivation-reoxygenation proximal tubular injury.

Abstract

The purpose of this study was to assess whether proximal renal tubules generate excess hydroxyl radical (.OH) during hypoxia/reoxygenation or ischemia/reperfusion injury, thereby supporting the hypothesis that reactive oxygen species contribute to the pathogenesis of postischemic acute renal failure. In the first phase of the study, rat isolated proximal tubular segments (PTS) were subjected to hypoxia (95% N2- 5% CO2) for 15, 30, or 45 min, followed by 15 to 30 min of reoxygenation in the presence of sodium salicylate, a stable .OH trap. Cellular injury after hypoxia and reoxygenation was assessed by lactate dehydrogenase release; .OH production was gauged by hydroxylated salicylate by-product generation (2,3-, 2,5-dihydroxybenzoic acids (DHBA); quantified by HPLC/electrochemical detection). Continuously oxygenated PTS served as controls. Despite substantial lactate dehydrogenase release during hypoxia (8 to 46%) and reoxygenation (8 to 11%), DHBA production did not exceed that of the coincubated, continuously oxygenated control PTS. In the second phase of the study, salicylate-treated rats were subjected to 25 or 40 min of renal arterial occlusion +/- 15 min of reperfusion. No increase in renal DHBA concentrations occurred during ischemia or reperfusion, compared with that in sham-operated controls. To validate the salicylate trap method, PTS were incubated with a known .OH-generating system (Fe2+/Fe3+); in addition, rats were treated with antioxidant interventions (oxypurinol plus dimethylthiourea). Fe caused marked DHBA production, and the antioxidants halved in vivo DHBA generation. In conclusion, these results suggest that exaggerated .OH production is not a consequence of O2 deprivation/reoxygenation tubular injury.

N-type calcium channel and renal injury

“…Remarkable increase of renal reactive oxygen spices (ROS) production has been reported in different hypertensive [ 66 – 69 ] and diabetic [ 70 – 73 ] animal models via a nicotinamide adenine dinucleotide phosphate (NADPH) oxidase-dependent manner [ 74 ]. Accumulating evidence from both in vivo and in vitro studies further elucidated specific contribution of ROS overproduction to renal injury by targeting different cell types: Oxidant-mediated injury to tubular cells was suggested to play a critical role in tubulointerstitial fibrosis [ 75 – 78 ]. High glucose induces proliferation and extracellular matrix (ECM) synthesis of mesangial cells through activating NADPH oxidase which thereby cause ROS production [ 79 – 81 ].…”