Background. Despite the beneficial effects of alagebrium (ALA), a putative advanced glycation end-product (AGE) breaker, on diabetic nephropathy, its renoprotective mechanisms are incompletely understood. Since oxidative stress exacerbates diabetic renal injury through interaction with AGE, the present study examined the antioxidative property of ALA in db/db mice, mesangial cells cultured under high glucose or H2O2 and a test tube.Methods. ALA (2 mg/kg/day) was administered intraperitoneally for 12 weeks to 8-week-old db/m and db/db (DALAE) mice or for 4 weeks to 16-week-old db/db mice (DALAL). Oxidative stress markers (nitrotyrosine accumulation, expression and translocation of nicotinamide adenine dinucleotide phosphate (NADPH) oxidase subunits, cellular DCF-DA fluorescence) together with urinary albumin excretion and histological changes including mesangial expansion were measured. The concentration of H2O2 in the presence and absence of ALA was measured by iodometric analysis in a test tube.Results. ALA significantly reduced not only urinary albumin excretion and renal pathological changes but also accumulation of pentosidine and nitrotyrosine and expression of NADPH oxidase subunits in db/db mice regardless of treatment protocol. In mesangial cells, ALA effectively prevented not only high glucose- but also H2O2-induced membrane translocation of NADPH oxidase subunit (p47 phox, p67 phox and rac1) and protein kinase C isoform (α, βI and βII) and Nox4 messenger RNA expression concomitant with cellular reactive oxygen species. Furthermore, ALA directly decreased H2O2 in a test tube.Conclusion. ALA has both direct and indirect antioxidant effects that may play important roles in ALA's renoprotective effect in diabetic kidneys.
Bibliographical noteFunding Information:
Acknowledgements. This work was supported, in part, by grant nos. R01-2006-000-10829, R15-2006-020, and R31-2008-000-10010-0 from the Korea Science and Engineering Foundation. P.J. was supported by RP-Grant 2009 (2009-1949-1-1) of Ewha Womans University and H.J.Y. by Brain Korea 21 project. ALA used in this study was kindly provided in 1999 by Peter Ulrich, PhD of the Kenneth S. Warren Laboratories, NY, USA and by Alteon Inc., Parsippany, NJ, USA in 2004 for additional studies.
- diabetic nephropathy
- reactive oxygen species