Research Article| Volume 192, ISSUE 1, P15-24, May 2007

Vascular NAD(P)H oxidase mediates endothelial dysfunction in basilar arteries from Otsuka Long-Evans Tokushima Fatty (OLETF) rats


      We examined the responses of basilar arteries taken from Otsuka Long-Evans Tokushima Fatty (OLETF) rats, a type 2 diabetes model. Both the nitric oxide (NO)-mediated relaxation and the cyclic 3′,5′-guanosine monophosphate (cGMP) production elicited by acetylcholine (ACh) were much weaker in OLETF rats than in age-matched control Long Evans Tokushima Otsuka (LETO) rats. The contraction induced by an NO synthase (NOS) inhibitor [NG-nitro-l-arginine (l-NNA)] was weaker in the OLETF group. In that group, application of apocynin, an NAD(P)H oxidase inhibitor, normalized (i) ACh-induced relaxation, (ii) l-NNA-induced contraction, and (iii) ACh-induced cGMP production to the LETO levels. Superoxide anion production was greater in basilar arteries from OLETF rats than in those from LETO rats. The protein expression of gp91phox, an NAD(P)H oxidase subunit, was upregulated in the OLETF arteries (versus LETO ones). These results suggest that the existence of endothelial dysfunction in basilar arteries in type 2 diabetes is related to increased oxidative stress mediated via NAD(P)H oxidase. Possibly, an impairment of NO-dependent relaxation responses and a basal impairment of NO signaling may be responsible for the increased risk of adverse cerebrovascular events in type 2 diabetes.


      To read this article in full you will need to make a payment

      Purchase one-time access:

      Academic & Personal: 24 hour online accessCorporate R&D Professionals: 24 hour online access
      One-time access price info
      • For academic or personal research use, select 'Academic and Personal'
      • For corporate R&D use, select 'Corporate R&D Professionals'


      Subscribe to Atherosclerosis
      Already a print subscriber? Claim online access
      Already an online subscriber? Sign in
      Institutional Access: Sign in to ScienceDirect


        • Faraci F.M.
        • Heistad D.D.
        Regulation of large cerebral arteries and cerebral microvascular pressure.
        Circ Res. 1990; 66: 8-17
        • Faraci F.M.
        • Heistad D.D.
        Regulation of the cerebral circulation: role of endothelium and potassium channels.
        Physiol Rev. 1998; 78: 53-97
        • Laakso M.
        • Lehto S.
        Epidemiology of risk factors for cardiovascular disease in diabetes and impaired glucose tolerance.
        Atherosclerosis. 1998; 137: S65-S73
        • De Vriese A.S.
        • Verbeuren T.J.
        • Van de Voorde J.
        • et al.
        Endothelial dysfunction in diabetes.
        Br J Pharmacol. 2000; 130: 963-974
        • Kobayashi T.
        • Taguchi K.
        • Yasuhiro T.
        • et al.
        Impairment of PI3-K/Akt pathway underlies attenuated endothelial function in aorta of type 2 diabetic mouse model.
        Hypertension. 2004; 44: 956-962
        • Kanie N.
        • Matsumoto T.
        • Kobayashi T.
        • et al.
        Relationship between peroxisome proliferator-activated receptors (PPAR alpha and PPAR gamma) and endothelium-dependent relaxation in streptozotocin-induced diabetic rats.
        Br J Pharmacol. 2003; 140: 23-32
        • Kobayashi T.
        • Kamata K.
        Effect of chronic insulin treatment on NO production and endothelium-dependent relaxation in aortae from established STZ-induced diabetic rats.
        Atherosclerosis. 2001; 155: 313-320
        • Matsumoto T.
        • Kobayashi T.
        • Kamata K.
        Alterations in EDHF-type relaxation and phosphodiesterase activity in mesenteric arteries from diabetic rats.
        Am J Physiol Heart Circ Physiol. 2003; 285: H283-H291
        • Pieper G.M.
        Review of alterations in endothelial nitric oxide production in diabetes: protective role of arginine on endothelial dysfunction.
        Hypertension. 1998; 31: 1047-1060
        • Kawano K.
        • Hirashima T.
        • Mori S.
        • et al.
        Spontaneous long-term hyperglycemic rat with diabetic complications. Otsuka Long-Evans Tokushima Fatty (OLETF) strain.
        Diabetes. 1992; 41: 1422-1428
        • Minami A.
        • Ishimura N.
        • Harada N.
        • et al.
        Exercise training improves acetylcholine-induced endothelium-dependent hyperpolarization in type 2 diabetic rats, Otsuka Long-Evans Tokushima Fatty rats.
        Atherosclerosis. 2002; 162: 85-92
        • Didion S.P.
        • Lynch C.M.
        • Baumbach G.L.
        • et al.
        Impaired endothelium-dependent responses and enhanced influence of Rho-kinase in cerebral arterioles in type II diabetes.
        Stroke. 2005; 36: 342-347
        • Erdos B.
        • Snipes J.A.
        • Miller A.W.
        • et al.
        Cerebrovascular dysfunction in Zucker obese rats is mediated by oxidative stress and protein kinase C.
        Diabetes. 2004; 53: 1352-1359
        • Schwaninger R.M.
        • Sun H.
        • Mayhan W.G.
        Impaired nitric oxide synthase-dependent dilatation of cerebral arterioles in type II diabetic rats.
        Life Sci. 2003; 73: 3415-3425
        • Matsumoto T.
        • Yoshiyama S.
        • Wakabayashi K.
        • et al.
        Effects of chronic insulin on endothelial dysfunction of basilar arteries from established streptozotocin-diabetic rats.
        Eur J Pharmacol. 2004; 504: 119-127
        • Faraci F.M.
        Oxidative stress: the curse that underlies cerebral vascular dysfunction?.
        Stroke. 2005; 36: 186-188
        • Cai H.
        • Griendling K.K.
        • Harrison D.G.
        The vascular NAD(P)H oxidases as therapeutic targets in cardiovascular diseases.
        Trends Pharmacol Sci. 2003; 24: 471-478
        • Hink U.
        • Li H.
        • Mollnau H.
        • et al.
        Mechanisms underlying endothelial dysfunction in diabetes mellitus.
        Circ Res. 2001; 88: e14-e22
        • Lassegue B.
        • Clempus R.E.
        Vascular NAD(P)H oxidases: specific features, expression, and regulation.
        Am J Physiol Regul Integr Comp Physiol. 2003; 285: R277-R297
        • Matsumoto T.
        • Sato A.
        • Suenaga H.
        • et al.
        Modulations of shear stress-induced contractile responses and agonist-induced vasodilation in hypercholesterolemic rats.
        Atherosclerosis. 2004; 175: 31-38
        • Wang H.D.
        • Pagano P.J.
        • Du Y.
        • et al.
        Superoxide anion from the adventitia of the rat thoracic aorta inactivates nitric oxide.
        Circ Res. 1998; 82: 810-818
        • Sugawara T.
        • Fujii S.
        • Zaman A.K.
        • et al.
        Coronary capillary network remodeling and hypofibrinolysis in aged obese diabetic rats: implications for increased myocardial vulnerability to ischemia.
        Mol Cell Biochem. 2003; 248: 165-170
        • Beckman J.S.
        • Chen J.
        • Ischiropoulos H.
        • et al.
        Oxidative chemistry of peroxynitrite.
        Methods Enzymol. 1994; 233: 229-240
        • Didion S.P.
        • Faraci F.M.
        Effects of NADH and NADPH on superoxide levels and cerebral vascular tone.
        Am J Physiol Heart Circ Physiol. 2002; 282: H688-H695
        • Ignarro L.J.
        • Fukuto J.M.
        • Griscavage J.M.
        • et al.
        Oxidation of nitric oxide in aqueous solution to nitrite but not nitrate: comparison with enzymatically formed nitric oxide from l-arginine.
        Proc Natl Acad Sci USA. 1993; 90: 8103-8107
        • Modin A.
        • Bjorne H.
        • Herulf M.
        • et al.
        Nitrite-derived nitric oxide: a possible mediator of ‘acidic-metabolic’ vasodilation.
        Acta Physiol Scand. 2001; 171: 9-16
        • Jackson T.S.
        • Xu A.
        • Vita J.A.
        • Keaney Jr, J.F.
        Ascorbate prevents the interaction of superoxide and nitric oxide only at very high physiological concentrations.
        Circ Res. 1998; 83: 916-922
        • Szocs K.
        • Lassegue B.
        • Sorescu D.
        • et al.
        Upregulation of Nox-based NAD(P)H oxidases in restenosis after carotid injury.
        Arterioscler Thoromb Vasc Biol. 2002; 22: 21-27
        • Wambi-Kiesse C.O.
        • Katusic Z.S.
        Inhibition of copper/zinc superoxide dismutase impairs NO-mediated endothelium-dependent relaxations.
        Am J Physiol Heart Circ Physiol. 1999; 276: H1043-H1048
        • Ago T.
        • Kitazono T.
        • Kuroda J.
        • et al.
        NAD(P)H oxidases in rat basilar arterial endothelial cells.
        Stroke. 2005; 36: 1040-1046
        • Sercombe R.
        • Vicaut E.
        • Oudart N.
        • et al.
        Acetylcholine-induced relaxation of rabbit basilar artery in vitro is rapidly reduced by reactive oxygen species in acute hyperglycemia: role of NADPH oxidase.
        J Cardiovasc Pharmacol. 2004; 44: 507-516
        • Inoguchi T.
        • Li P.
        • Umeda F.
        • et al.
        High glucose level and free fatty acid stimulate reactive oxygen species production through protein kinase C-dependent activation of NAD(P)H oxidase in cultured vascular cells.
        Diabetes. 2000; 49: 1939-1945
        • Sonta T.
        • Inoguchi T.
        • Tsubouchi H.
        • et al.
        Evidence for contribution of vascular NAD(P)H oxidase to increased oxidative stress in animal models of diabetes and obesity.
        Free Radic Biol Med. 2004; 37: 115-123
        • Phillips S.A.
        • Sylvester F.A.
        • Frisbee J.C.
        Oxidative stress and constrictor reactivity impair cerebral artery dilation in obese Zucker rats.
        Am J Physiol Regul Integr Comp Physiol. 2005; 288: R522-R530