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(Arteriosclerosis, Thrombosis, and Vascular Biology. 2002;22:2037.)
© 2002 American Heart Association, Inc.

Atherosclerosis

Cytochrome b₅₅₈–Dependent NAD(P)H Oxidase–Phox Units in Smooth Muscle and Macrophages of Atherosclerotic Lesions

Natalia Kalinina; Alex Agrotis; Eduard Tararak; Yulia Antropova; Peter Kanellakis; Olga Ilyinskaya; Mark T. Quinn; Vladimir Smirnov; Alex Bobik

From the Baker Medical Research Institute (N.K., A.A., P.K., A.B.), Alfred Hospital, Melbourne, Australia; the Institute of Experimental Cardiology (N.K., E.T., Y.A., O.I., V.S.), Cardiology Research Center, Moscow, Russia; and the Department of Veterinary Molecular Biology (M.T.Q.), Montana State University, Bozeman.

Correspondence to Dr Natalia Kalinina, Institute of Experimental Cardiology, Cardiology Research Center, Moscow 121552, Russia. E-mail cardiocell{at}cardio.ru

Abstract

Objective— Despite studies implicating superoxide anion–producing oxidases in atherosclerosis, their characteristics, expression, and regulation in cells of lesions are poorly understood. We examined the following: (1) whether cytochrome b₅₅₈–dependent NAD(P)H oxidase–phox peptides are expressed by intimal smooth muscle cells (iSMCs) and macrophages of human aortic atherosclerotic lesions and their regulation and (2) whether cytochrome b₅₅₈–dependent NAD(P)H oxidase represents a major NAD(P)H oxidase in iSMCs.

Methods and Results— Using a combination of immunochemical and reverse transcription–polymerase chain reaction procedures, we demonstrate that p22^phox and gp91^phox (cytochrome b₅₅₈) expression in normal intima was restricted to a quarter of the iSMCs. In fatty streaks, a similar fraction of iSMCs expressed cytochrome b₅₅₈, whereas macrophages also expressed low levels of p47^phox and p67^phox. In fibrofatty lesions, the majority of iSMCs expressed the cytochrome b₅₅₈ subunits; p67^phox was also detected. Macrophages and macrophage-derived foam cells expressed the 4 phox subunits that constitute superoxide-producing cytochrome b₅₅₈–dependent NAD(P)H oxidase. These were upregulated by transforming growth factor-ß₁ and interferon-. Aortic lesions also expressed Thox1 and Nox4, and although their expression also increases with lesion severity, their expression is less frequent than that of gp91^phox.

Conclusions— In human aortic fibrofatty lesions, a cytochrome b₅₅₈–dependent NAD(P)H oxidase appears to be a major iSMC and macrophage oxidase whose expression is upregulated by cytokines.

Key Words: atherosclerosis • gp91^phox • Thox1 • Nox4 • cytokines

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