title: [Role of NADPH oxidase in oxidative stress injury of human dermal fibroblasts].
abstract: OBJECTIVE
  To investigate the role of NADPH oxidase (Nox) in the oxidative stress injury of human dermal fibroblasts (HFbs).
  
  
  METHODS
  An oxidative stress injury model was established in HFbs by exposure to H(2)O(2). Normal HFbs and HFbs exposed to H(2)O(2) with and without pretreatment with NADPH oxidase inhibitor were tested for cell viability using MTT assay, and the intracellular reactive oxygen species (ROS) were determined with a DCFH-DA fluorescent probe. Western blotting was used to measure the protein expressions of membrane-bound subunit gp91phox of NADPH oxidase in the cells.
  
  
  RESULTS
  H(2)O(2) time- and concentration-dependently induced oxidative stress injury in the fibroblasts, causing a reduction of the cell viability to 40% after a 24-h exposure at 700 µmol/L (P<0.05) and an increase of ROS by 2 folds after a 2-h exposure at 700 µmol/L (P<0.05). Compared with the cells with oxidative stress injury, the cells with NADPH oxidase inhibitor pretreatment showed a 20% higher cell viability (P<0.05) and normal ROS level (P<0.05) following H(2)O(2) exposure. Western blotting demonstrated increased expression of gp91phox in the cells exposed to increasing H(2)O(2) concentrations, but gp91phox expression remained normal in cells pretreated with NADPH oxidase inhibitor.
  
  
  CONCLUSIONS
  H(2)O(2) can induce oxidative stress injury in the fibroblasts by affecting NADPH oxidase, especially its membrane-bound subunit gp91phox.
authors.affiliation: Department of Dermatology, Shunde First People's Hospital Affiliated to Southern Medical University, FoShan 528300, China.E-mail: chenying_chun@163.com.
authors.forename: Ying
authors.initials: Y
authors.lastname: Chen
authors.forename: Hao
authors.initials: H
authors.lastname: Huang
authors.forename: Hong-Feng
authors.initials: HF
authors.lastname: Tang
authors.forename: Xiu-Fen
authors.initials: XF
authors.lastname: Zheng
authors.forename: Yong
authors.initials: Y
authors.lastname: Hu
authors.forename: Rui-Hua
authors.initials: RH
authors.lastname: Wang
mesh_terms: D002470:Cell Survival; D002478:Cells, Cultured; D005347:Fibroblasts / Q000166:cytology* / Q000201:enzymology; D006801:Humans; D006861:Hydrogen Peroxide; D008562:Membrane Glycoproteins / Q000378:metabolism; D000074662:NADPH Oxidase 2; D019255:NADPH Oxidases / Q000037:antagonists & inhibitors / Q000378:metabolism*; D010084:Oxidation-Reduction; D018384:Oxidative Stress; D017382:Reactive Oxygen Species / Q000378:metabolism
chemical_list: D008562:Membrane Glycoproteins; D017382:Reactive Oxygen Species; D006861:Hydrogen Peroxide; C056558:CYBB protein, human; D000074662:NADPH Oxidase 2; D019255:NADPH Oxidases