Endothelial-derived nitric oxide preserves anticoagulant heparan sulfate expression in cultured porcine aortic endothelial cells

Masahiko Irokawa, Masanori Nishinaga, Uichi Ikeda, Yuichi Shinoda, Makoto Suematsu, Nobuhito Gouda, Yuzuru Ishimura, Kazuyuki Shimada

Research output: Contribution to journalArticle

12 Citations (Scopus)

Abstract

Nitric oxide (NO) has been shown to inhibit platelet adhesion and aggregation, but there are no reports on its interaction with the coagulation system. We investigated the effect of the L-arginine analogues, N-nitro-L- arginine (LNA), N(G)-nitro-L-arginine methyl ester (L-NAME), and N(G)- monomethyl-L arginine (L-NMMA), competitive inhibitors of NO production, on endothelial-surface heparan sulfate. Addition of LNA to porcine aortic endothelial cells reduced 125I-labeled antithrombin III binding to the cell surface heparan sulfate in a dose- and time-dependent fashion. Significant inhibition was observed with 1 mM LNA, and the maximal suppression (- 50% of control) occurred at 10 mM LNA after 12 h. L-NAME (1 mM) and L-NMMA (1 mM) also significantly inhibited the antithrombin III binding. The iron chelator desferrioxamine significantly prevented the reduction of antithrombin III binding to LNA-treated cells. We further investigated the effect of L-NAME on intracellular oxidative stress of endothelial cells using a hydroperoxide-sensitive fluorochrome, carboxy- dichloro-dihydrofluorescein diacetate bisacetoxymethyl ester probe, and revealed that inhibition of NO synthesis by L-NAME led to a marked increase in intracellular oxidative stress. These results demonstrated that the prolonged inhibition of NO synthesis in porcine aortic endothelial cells decreases the expression of anticoagulant heparan sulfate on endothelial cells through the increase in intracellular oxidative stress, perhaps comprising another mechanism by which NO affects the coagulation system in the vasculature.

Original languageEnglish
Pages (from-to)9-17
Number of pages9
JournalAtherosclerosis
Volume135
Issue number1
DOIs
Publication statusPublished - 1997 Nov

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Heparitin Sulfate
NG-Nitroarginine Methyl Ester
Anticoagulants
Nitric Oxide
Swine
Endothelial Cells
Antithrombin III
Arginine
omega-N-Methylarginine
Oxidative Stress
Deferoxamine
Chelating Agents
Platelet Aggregation
Fluorescent Dyes
Hydrogen Peroxide
Esters
Iron

Keywords

  • Antithrombin III
  • EDRF
  • Oxidative stress
  • Proteoglycan
  • Thrombosis

ASJC Scopus subject areas

  • Cardiology and Cardiovascular Medicine

Cite this

Endothelial-derived nitric oxide preserves anticoagulant heparan sulfate expression in cultured porcine aortic endothelial cells. / Irokawa, Masahiko; Nishinaga, Masanori; Ikeda, Uichi; Shinoda, Yuichi; Suematsu, Makoto; Gouda, Nobuhito; Ishimura, Yuzuru; Shimada, Kazuyuki.

In: Atherosclerosis, Vol. 135, No. 1, 11.1997, p. 9-17.

Research output: Contribution to journalArticle

Irokawa, M, Nishinaga, M, Ikeda, U, Shinoda, Y, Suematsu, M, Gouda, N, Ishimura, Y & Shimada, K 1997, 'Endothelial-derived nitric oxide preserves anticoagulant heparan sulfate expression in cultured porcine aortic endothelial cells', Atherosclerosis, vol. 135, no. 1, pp. 9-17. https://doi.org/10.1016/S0021-9150(97)00117-2
Irokawa, Masahiko ; Nishinaga, Masanori ; Ikeda, Uichi ; Shinoda, Yuichi ; Suematsu, Makoto ; Gouda, Nobuhito ; Ishimura, Yuzuru ; Shimada, Kazuyuki. / Endothelial-derived nitric oxide preserves anticoagulant heparan sulfate expression in cultured porcine aortic endothelial cells. In: Atherosclerosis. 1997 ; Vol. 135, No. 1. pp. 9-17.
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