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The roles of NADPH‐oxidase and nNOS for the increased oxidative stress and the oxygen consumption in the diabetic kidney
Author(s) -
Edlund Jenny,
Fasching Angelica,
Liss Per,
Hansell Peter,
Palm Fredrik
Publication year - 2010
Publication title -
diabetes/metabolism research and reviews
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.307
H-Index - 110
eISSN - 1520-7560
pISSN - 1520-7552
DOI - 10.1002/dmrr.1099
Subject(s) - apocynin , nadph oxidase , nad(p)h oxidase , nicotinamide adenine dinucleotide phosphate , endocrinology , oxidative stress , medicine , chemistry , reactive oxygen species , kidney , oxygen , diabetes mellitus , nitric oxide , nitric oxide synthase , oxidase test , biochemistry , biology , enzyme , organic chemistry
Abstract Background Sustained hyperglycaemia induces increased renal oxygen consumption resulting in reduced oxygen availability in the diabetic kidney. We investigated the roles of the nicotinamide adenine dinucleotide phosphate (NADPH)‐oxidase and the neuronal nitric oxide synthase (nNOS) for the increased oxygen consumption in streptozotocin‐diabetic rats. Methods Oxygen consumption was measured in isolated proximal tubular cells (PTC) from streptozotocin‐induced diabetic rats ( n = 7–9 per group) with and without chronic treatment with apocynin, a NADPH‐oxidase inhibitor, or S ‐methyl‐ L ‐thiocitrulline (SMTC), a selective nNOS inhibitor, or a combination of the two and the results were compared to normoglycaemic controls ( n = 10). Oxidative stress was estimated from thiobarbituric acid reactive substances and protein expression measured by Western blot. Results Proximal tubular cells from untreated diabetic rats had increased oxygen consumption compared to controls (40.6 ± 7.9 versus 10.9 ± 2.0 nmol/mg protein/min). All treatments reduced the diabetes‐induced increase in oxygen consumption (apocynin 10.5 ± 1.7, SMTC 19.7 ± 3.0 and apocynin + SMTC 21.6 ± 3.6 nmol/mg protein/min). Neither apocynin nor SMTC had any effect on the oxygen consumption in cells pre‐incubated with ouabain, an inhibitor of active electrolyte transport. Oxidative stress was elevated in the diabetic kidney and inhibited by all treatments. The increased oxygen consumption by diabetic proximal tubular cells correlated with increased protein expressions of p47 phox and nNOS and the treatments prevented these increases. Conclusions Diabetes induces oxidative stress, which increases oxygen consumption in proximal tubular cells. Inhibition of either NADPH‐oxidase or nNOS prevented the increased oxygen consumption. The effect of blocking both these enzymes was less than additive suggesting overlapping pathways which warrant further studies. Copyright © 2010 John Wiley & Sons, Ltd.