title: Cigarette smoke particle-phase extract induces HO-1 expression in human tracheal smooth muscle cells: role of the c-Src/NADPH oxidase/MAPK/Nrf2 signaling pathway.
abstract: Heme oxygenase-1 (HO-1) is known as an oxidative stress protein that is up-regulated by various stimuli. HO-1 has been shown to protect cells against oxidative damage. Cigarette smoke is a potential inflammatory mediator that causes chronic obstructive pulmonary disease and asthma. In this study, we report that cigarette smoke particle-phase extract (CSPE) is an inducer of HO-1 expression mediated through various signaling pathways in human tracheal smooth muscle cells (HTSMCs). CSPE-induced HO-1 protein, mRNA expression, and promoter activity were attenuated by pretreatment with a ROS scavenger (N-acetyl-l-cysteine) and inhibitors of c-Src (PP1), NADPH oxidase [diphenylene iodonium chloride (DPI) and apocynin (APO)], MEK1/2 (U0126), p38 MAPK (SB202190), and JNK1/2 (SP600125) or transfection with siRNAs for Src, p47(phox), NOX2, p42, p38, JNK2, or NF-E2-related factor 2 (Nrf2). CSPE-stimulated translocation of p47(phox) and Nrf2, ROS production, and NADPH oxidase activity was attenuated by transfection with siRNAs for Src, p47(phox), and NOX2 or pretreatment with PP1, DPI, or APO. Furthermore, CSPE-induced NOX2, c-Src, and p47(phox) complex formation was revealed by immunoprecipitation using an anti-NOX2, anti-p47(phox), or anti-c-Src Ab followed by Western blot against anti-NOX2, anti-p47(phox), or anti-c-Src Abs. These results demonstrate that CSPE-induced ROS generation is mediated through a c-Src/NADPH oxidase/MAPK pathway and in turn initiates the activation of Nrf2 and ultimately induces HO-1 expression in HTSMCs.
authors.affiliation: Department of Physiology and Pharmacology, Chang Gung University, Kwei-San, Tao-Yuan 33302, Taiwan.
authors.forename: Shin-Ei
authors.initials: SE
authors.lastname: Cheng
authors.forename: I-Ta
authors.initials: IT
authors.lastname: Lee
authors.forename: Chih-Chung
authors.initials: CC
authors.lastname: Lin
authors.forename: Yu Ru
authors.initials: YR
authors.lastname: Kou
authors.forename: Chuen-Mao
authors.initials: CM
authors.lastname: Yang
mesh_terms: D000111:Acetylcysteine / Q000494:pharmacology; D002478:Cells, Cultured; D045424:Complex Mixtures / Q000008:administration & dosage / Q000009:adverse effects*; D004791:Enzyme Inhibitors / Q000494:pharmacology; D051547:Heme Oxygenase-1 / Q000096:biosynthesis* / Q000235:genetics; D006801:Humans; D020929:Mitogen-Activated Protein Kinase Kinases / Q000235:genetics / Q000378:metabolism; D032389:Myocytes, Smooth Muscle / Q000187:drug effects / Q000378:metabolism* / Q000473:pathology; D019255:NADPH Oxidases / Q000037:antagonists & inhibitors / Q000096:biosynthesis / Q000235:genetics; D051267:NF-E2-Related Factor 2 / Q000235:genetics / Q000378:metabolism; D018384:Oxidative Stress / Q000187:drug effects; D052638:Particulate Matter / Q000009:adverse effects*; D044367:Phase Transition; D034741:RNA, Small Interfering / Q000235:genetics; D015398:Signal Transduction / Q000187:drug effects; D012907:Smoking / Q000009:adverse effects* / Q000378:metabolism; D014132:Trachea / Q000473:pathology; D019061:src-Family Kinases / Q000037:antagonists & inhibitors / Q000096:biosynthesis / Q000235:genetics
chemical_list: D045424:Complex Mixtures; D004791:Enzyme Inhibitors; D051267:NF-E2-Related Factor 2; D052638:Particulate Matter; D034741:RNA, Small Interfering; D051547:Heme Oxygenase-1; D019255:NADPH Oxidases; D019061:src-Family Kinases; D020929:Mitogen-Activated Protein Kinase Kinases; D000111:Acetylcysteine