Vascular NADH Oxidase Is Involved in Impaired Endothelium-Dependent Vasodilation in OLETF Rats, a Model of Type 2 Diabetes

Vascular NADH Oxidase Is Involved in Impaired Endothelium-Dependent Vasodilation in OLETF Rats, a Model of Type 2 Diabetes Yong K. Kim , Mi-S. Lee , Seok M. Son , In J. Kim , Won S. Lee , Byung Y. Rhim , Ki W. Hong and Chi D. Kim From the Department of Internal Medicine and Pharmacology, College of Medicine, Pusan National University, Pusan, South Korea Abstract Superoxide anion can modulate vascular smooth muscle tone and is potentially involved in diabetic vascular complications. The present study was undertaken to characterize both vascular production and the enzymatic source of superoxide anion in type 2 diabetic rats. In the thoracic aorta of OLETF rats, endothelium-dependent relaxation was markedly attenuated compared with that of control (LETO) rats in association with a significant increase in superoxide production (2,421.39 ± 407.01 nmol · min −1 · mg −1 ). The increased production of superoxide anion was significantly attenuated by diphenyleneiodonium (DPI; 10 μmol/l), an inhibitor of NAD(P)H oxidase. The production of superoxide anion in response to NADH as a substrate was markedly increased in the vascular homogenates, but NADPH, arachidonic acid, xanthine, and succinate produced only small increases in chemiluminescence. In line with these results, studies using various enzyme inhibitors, such as DPI, allopurinol, rotenone, N G -monomethyl- l -arginine, and indomethacin, suggest that the predominant source of superoxide anion in vascular particulate fraction is NADH-dependent membrane-bound oxidase. Furthermore, the expression of p22phox, a major component of vascular NAD(P)H oxidase, was markedly increased in the aorta from OLETF rats compared with that of LETO rats. These findings suggest that upregulated expression of p22phox mRNA and enhanced NADH oxidase activity contribute to the impaired endothelium-dependent vasodilation in OLETF rats. Footnotes Address correspondence and reprint requests to Chi D. Kim, Department of Pharmacology, College of Medicine, Pusan National University, Ami-dong 1-Ga, Seo-gu, Busan 602–739, Korea. E-mail: chidkim{at}pusan.ac.kr . Received for publication 3 April 2001 and accepted in revised form 5 November 2001. ACh, acetylcholine; DPI, diphenyleneiodonium; eNOS, endothelial nitric oxide synthase; l -NMMA, N G -monomethyl- l -arginine; NO, nitric oxide; ROS, reactive oxygen species; SNP, sodium nitroprusside; SOD, superoxide dismutase.