title: Regulation of NADPH oxidase subunit p22(phox) by NF-kB in human aortic smooth muscle cells.
abstract: Accumulating evidence demonstrates the involvement of oxidative stress in the pathophysiology of cardiovascular diseases. The molecular mechanisms accountable for the increased production of reactive oxygen species remain uncertain. Among others, NADPH oxidase is one of the most important sources of superoxide in vascular cells. Here we investigate the role of NF-kB in the regulation of p22(phox) subunit and NADPH oxidase activity, in human aortic smooth muscle cells. Overexpression of p65/RelA or IKKbeta up-regulated p22(phox) gene promoter activity. Transcription factor pull-down assays demonstrated the physical interaction of p65/RelA protein with predicted NF-kB binding sites. Real time PCR and Western blotting analysis showed that p22(phox) mRNA and protein expression are significantly down-regulated by NF-kB decoy oligodeoxynucleotides and N-alpha-tosyl-l-phenylalanine chloromethyl ketone (TPCK). Lucigenin-enhanced chemiluminescence assay revealed that NF-kB inhibitors reduce the NADPH-dependent superoxide production. Regulation of NADPH oxidase by NF-kB may represent a possible mechanism whereby pro-inflammatory factors induce oxidative stress in atherosclerosis, hypertension, diabetes, stroke or heart failure.
authors.affiliation: Nicolae Simionescu Institute of Cellular Biology and Pathology, 8 B.P. Hadeu Street, Bucharest, Romania.
authors.forename: A
authors.initials: A
authors.lastname: Manea
authors.forename: S A
authors.initials: SA
authors.lastname: Manea
authors.forename: A V
authors.initials: AV
authors.lastname: Gafencu
authors.forename: M
authors.initials: M
authors.lastname: Raicu
mesh_terms: D001011:Aorta / Q000166:cytology*; D001483:Base Sequence; D001665:Binding Sites; D002470:Cell Survival; D006801:Humans; D032389:Myocytes, Smooth Muscle / Q000378:metabolism*; D019255:NADPH Oxidases / Q000235:genetics / Q000378:metabolism*; D016328:NF-kappa B / Q000235:genetics / Q000378:metabolism*; D009841:Oligonucleotides / Q000235:genetics; D011401:Promoter Regions, Genetic / Q000235:genetics; D012333:RNA, Messenger / Q000235:genetics; D013481:Superoxides / Q000378:metabolism; D014162:Transfection; D015854:Up-Regulation
chemical_list: D016328:NF-kappa B; D009841:Oligonucleotides; D012333:RNA, Messenger; D013481:Superoxides; D019255:NADPH Oxidases