Advertisement

High glucose induced NF-κB DNA-binding activity in HAEC is maintained under low shear stress but inhibited under high shear stress: role of nitric oxide

      Abstract

      In the present study, we investigated whether low shear (LS, 2 dyn/cm2) favors high glucose (HG, 30 mM) induced nuclear factor kappa B (NF-κB) activity by regulating NO release in human aortic endothelial cells (HAEC). The results show that (i) under LS, the NF-κB activity of HAEC exposed to HG was significantly higher than HAEC in normal glucose (NG, 5.5 mM) (P<0.05). In contrast, under HS, the activation of NF-κB in HAEC exposed to HG showed no significant difference compared to that of NG. (ii) The NF-κB activity induced by HG is suppressed by high shear (HS) in the absence of a NO synthase inhibitor, -nitro-l-arginine methyl ester (l-NAME) but restored in its presence, while LS+HG induced NF-κB activity remains the same in the presence or absence of l-NAME. (iii) Endothelial nitric oxide synthase (eNOS) protein expression and quantitative detection of NO indicated that high shear stress significantly induced higher eNOS expression and NO production compared to low shear stress condition. Collectively, these data suggest that HS exerts a protective effect on HG induced NF-κB activity through NO mediated signaling. LS, on the other hand, may down-regulate eNOS expression resulting in reduced NO release, and thereby maintain high glucose induced NF-κB DNA-binding activity. These observations explain, in part, the mechanism by means of which hyperglycemia accelerates the focal development of atherosclerotic lesions in low shear (lesion prone) areas of the arterial tree.

      Keywords

      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:

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

      References

        • Feener E.P
        • King G.L
        Vascular dysfunction in diabetes mellitus.
        Lancet. 1997; 350: SI9-S13
        • Asakura T
        • Karino T
        Flow patterns and spatial distribution of atherosclerotic lesions in human coronary arteries.
        Circ. Res. 1990; 66: 1045-1066
      1. Hajra L, Evans AI, Chen M, Hyduk SJ, Collins T, Cybulsky MI. The NF-kappa B signal transduction pathway in aortic endothelial cells is primed for activation in regions predisposed to atherosclerotic lesion formation, vol. 97. PNAS 2000; 97: 9052–57.

        • Berliner J.A
        • Navab M
        • Fogelman A.M
        • Frank J.S
        • Demer L.L
        • Edwards P.A
        • et al.
        Atherosclerosis: basic mechanisms, oxidation.
        Inflamm. Genet. 1995; 91: 2488-2496
        • Mohan S
        • Mohan N
        • Sprague E.A
        Differential activation of NF-kappa B in human aortic endothelial cells conditioned to specific flow environments.
        Am. J. Physiol. 1997; 273: C572-C578
        • Ghosh S
        • May M.J
        • Kopp E.B
        NF-kappa B and Rel proteins: evolutionarily conserved mediators of immune responses.
        Annu. Rev. Immunol. 1998; 16: 225-260
        • Verma I.M
        • Stevenson J.K
        • Schwarz E.M
        • Van Antwerp D
        • Miyamoto S
        Rel/NF-kappa B/I kappa B family: Intimate tales of association and dissociation.
        Genes Develop. 1995; 9: 2723-2735
        • Pieper G.M
        • Riazul H
        Activation of nuclear factor-kappa B in cultured endothelial cells by increased glucose concentration: prevention by calphostin C.
        J. Cardiovasc. Pharmacol. 1997; 30: 528-532
        • Bao X
        • Lu C
        • Frangos J.A
        Temporal gradient in shear but not steady shear stress induces PDGFA and MCP-1 expression in endothelial cells: role of NO, NF kappa B, and egr-1.
        Arterioscler. Thromb. Vasc. Biol. 1999; 19: 996-1003
        • Nagel T
        • Resnick N
        • Dewey Jr., C.F
        • Gimbrone Jr., M.A
        Vascular endothelial cells respond to spatial gradients in fluid shear stress by enhanced activation of transcription factors.
        Arterioscler. Thromb. Vasc. Biol. 1999; 19: 1825-1834
        • Mohan S
        • Mohan N
        • Valente A.J
        • Sprague E.A
        Regulation of low shear flow-induced HAEC VCAM-1 expression and monocyte adhesion.
        Am. J. Physiol. 1999; 276: C1100-C1107
        • Boo Y.C
        • Sorescu G
        • Boyd N
        • Shiojima I
        • Walsh K
        • Du J
        • et al.
        Shear stress stimulates phosphorylation of endothelial nitric oxide synthase at Ser 1179 by Akt-independent mechanisms: role of protein kinase A.
        J. Biol. Chem. 2002; 277: 3388-3396
        • Hamuro M
        • Polan J
        • Natarajan M
        • Mohan S
        High glucose induced nuclear factor kappa B mediated inhibition of endothelial cell migration.
        Atherosclerosis. 2002; 162: 277-287
      2. Xiao Z, Zhang Z, Ranjan V, Diamond SL. Shear stress induction of the endothelial nitric oxide synthase gene is calcium-dependent but not calcium-activated. J Cell Physiol 1997;171:205–11 [published Erratum appeared in J Cell Physiol Jan 1998;174(1):144].

        • Du X.L
        • Edelstein D
        • Dimmeler S
        • Ju Q
        • Sui C
        • Brownlee M
        Hyperglycemia inhibits endothelial nitric oxide synthase activity by post-translational modification at the Akt site.
        J. Clin. Invest. 2001; 108: 1341-1348
        • Ming-Hui Z
        • Chaomei S
        • Richard A.C
        High glucose via peroxynitrite causes tyrosine nitration and inactivation of prostacyclin synthase that is associated with thromboxane/prostaglandin H2 receptor mediated apoptosis and adhesion molecule expression in cultured human aortic endothelial cells.
        Diabetes. 2002; 51: 198-203
        • Inoguchi T
        • Li P
        • Umeda F
        • Yu H.Y
        • Kakimot M
        • Imamura M
        • 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
        • Chakravarthy U
        • Hayes R.G
        • Stitt A.W
        • McAuley E
        • Archer D.B
        Constitutive nitric oxide synthase expression in retinal vascular endothelial cells is suppressed by high glucose and advanced glycation end products.
        Diabetes. 1998; 47: 945-952
        • Noyman I
        • Marikovsky M
        • Sasson S
        • Stark A.H
        • Bernath K
        • Seger R
        • et al.
        Hyperglycemia reduces nitric oxide synthase and glycogen synthase activity in endothelial cells.
        Nitric oxide. 2002; 7: 187-193
      3. Malek AM, Izumo S, Alper SL. Modulation by pathophysiological stimuli of the shear stress-induced up-regulation of endothelial nitric oxide synthase expression in endothelial cells. Neurosurgery 1999;45:334–45.

        • Ranjan V
        • Xiao Z
        • Diamond S.L
        Constitutive NOS expression in cultured endothelial cells is elevated by fluid shear stress.
        Am. J. Physiol. 1995; 269: H550-H555
        • Ziegler T
        • Silacci P
        • Harrison V.J
        • Hayoz D
        Nitric oxide synthase expression in endothelial cells exposed to mechanical forces.
        Hypertension. 1998; 32: 351-355
        • Channon K.M
        • Qian H
        • George S.E
        Nitric oxide synthase in atherosclerosis and vascular injury: insights from experimental gene therapy.
        Arterioscler. Thromb. Vasc. Biol. 2000; 20: 1873-1881
        • Dimmeler S
        • Fleming I
        • Fisslthaler B
        • Hermann C
        • Busse R
        • Zeiher A.M
        Activation of nitric oxide synthase in endothelial cells by Akt-dependent phosphorylation.
        Nature. 1999; 399: 601-605
        • Spiecker M
        • Peng H.B
        • Liao J.K
        Inhibition of endothelial vascular cell adhesion molecule-1 expression by nitric oxide involves the induction and nuclear translocation of IkappaB-alpha.
        J. Biol. Chem. 1997; 272: 30969-30974
        • Peng H.B
        • Libby P
        • Liao J.K
        Induction and stabilization of I kappa B alpha by nitric oxide mediates inhibition of NF-kappa B.
        J. Biol. Chem. 1995; 270: 14214-14219
        • DelaTorre A
        • Schroeder R.A
        • Punzalan C
        • Kuo P.C
        Endotoxin-mediated S-nitrosylation of p50 alters NF-kappa B-dependent gene transcription in ANA-1 murine macrophages.
        J. Immunol. 1999; 162: 4101-4108
        • Matthews J.R
        • Botting C.H
        • Panico M
        • Morris H.R
        • Hay R.T
        Inhibition of NF-kappaB DNA binding by nitric oxide.
        Nucl. Acid Res. 1996; 24: 236-2242
        • Wei Z
        • Costa K
        • Al-Mehdi A.B
        • Dodia C
        • Muzykanto V
        • Fisher A.B
        Simulated Ischemia in flow-adapted endothelial cells leads to generation of reactive oxygen species and cell signaling.
        Circ Res. 1999; 85: 682-689