p22phox is a critical component of the superoxide-generating NADH/NADPH oxidase system and regulates angiotensin II-induced hypertrophy in vascular smooth muscle cells.
Superoxide anion formation is vital to the microbicidal activity of phagocytes. Recently, however, there is accumulating evidence that it is also involved in cell growth in vascular smooth muscle cells (VSMCs). We have shown that the hypertrophic agent angiotensin II stimulates superoxide production by activating the membrane-bound NADH/NADPH oxidase and that inhibition of this oxidase attenuates vascular hypertrophy. However, the molecular identity of this oxidase in VSMCs is unknown. We have recently cloned the cytochrome b558 alpha-subunit, p22(phox) (one of the key electron transfer elements of the NADPH oxidase in phagocytes), from a rat VSMC cDNA library, but its role in VSMC oxidase activity remains unclarified. Here we report that the complete inhibition of p22(phox) mRNA expression by stable transfection of antisense p22(phox) cDNA into VSMCs results in a decrease in cytochrome b content, which is accompanied by a significant inhibition of angiotensin II-stimulated NADH/NADPH-dependent superoxide production, subsequent hydrogen peroxide production, and [3H]leucine incorporation. We provide the first evidence that p22(phox) is a critical component of superoxide-generating vascular NADH/NADPH oxidase and suggest a central role for this oxidase system in vascular hypertrophy.
Ushio-Fukai
M
M
Department of Medicine, Division of Cardiology, Emory University, Atlanta, Georgia 30322, USA.
Zafari
A M
AM
Fukui
T
T
Ishizaka
N
N
Griendling
K K
KK
D000804:Angiotensin II / Q000494:pharmacology*; D000818:Animals; D000959:Antihypertensive Agents / Q000494:pharmacology; D001713:Biphenyl Compounds / Q000494:pharmacology; D001808:Blood Vessels / Q000254:growth & development; D003573:Cytochrome b Group / Q000096:biosynthesis; D016373:DNA, Antisense / Q000235:genetics; D018076:DNA, Complementary / Q000235:genetics; D006861:Hydrogen Peroxide / Q000378:metabolism; D006984:Hypertrophy / Q000139:chemically induced; D007093:Imidazoles / Q000494:pharmacology; D019808:Losartan; D008297:Male; D026901:Membrane Transport Proteins; D009131:Muscle, Smooth, Vascular / Q000201:enzymology* / Q000473:pathology; D009247:NADH, NADPH Oxidoreductases / Q000378:metabolism*; D009252:NADPH Dehydrogenase / Q000235:genetics / Q000378:metabolism*; D019255:NADPH Oxidases; D010750:Phosphoproteins / Q000235:genetics / Q000378:metabolism*; D051381:Rats; D017207:Rats, Sprague-Dawley; D015398:Signal Transduction; D013481:Superoxides / Q000378:metabolism*; D013777:Tetrazoles / Q000494:pharmacology; D014162:Transfection; D014662:Vasoconstrictor Agents / Q000494:pharmacology*
D000959:Antihypertensive Agents; D001713:Biphenyl Compounds; D003573:Cytochrome b Group; D016373:DNA, Antisense; D018076:DNA, Complementary; D007093:Imidazoles; D026901:Membrane Transport Proteins; D010750:Phosphoproteins; D013777:Tetrazoles; D014662:Vasoconstrictor Agents; D013481:Superoxides; D000804:Angiotensin II; D006861:Hydrogen Peroxide; D009247:NADH, NADPH Oxidoreductases; D019255:NADPH Oxidases; C086986:CYBA protein, human; D009252:NADPH Dehydrogenase; D019808:Losartan