title: [Expression and significance of Nrf2/ARE pathway ralated factors in the HepG2 cell model of steatosis].
abstract: OBJECTIVE
  To explore a new method of establishing HepG2 cell model of steatosis and observe the expression and significance of nuclear factor erythroid-2p45-related factor 2(Nrf2)/antioxidative response element (ARE) pathway related factors in HepG2 cells of steatosis.
  
  
  METHODS
  HepG2 cells were induced with DMEM containing 25% fetal bovine serum, 0.1% MCT/LCT Fat Emulsion and 0.1 mmol/L free fatty acid (FFA) at different stages and the control group cells were cultured with normal DMEM medium. After the cell models were successfully established, lipid droplets in cytoplasm were observed with Oil Red 0 staining, and the triglyceride (TG) accumulation in HepG2 cells were tested by biochemical assay. Intracellular reactive oxygen species (ROS) concentration were detected by flow cytometry. Nitric oxide (NO), superoxide dismutase(SOD), malonyldialdehyde(MDA) and glutathione peroxidase(GSH-Px) were tested by biological reagent kit, while the protein expression of nuclear factor erythroid-2p45-related factor 2(Nrf2), heme oxygenase-1 (HO-1) and
  quinone oxidoreductase-1(NQO1) were analyzed by Western blot.
  
  
  RESULTS
  Compared with that in the control group, red cytoplasmic lipid droplets were visible in model group; TG,ROS, NO, MDA concentration (P < 0.05, P < 0.01) and the protein expression of Nrf2, HO-1 and NQO1 (P < 0.05, P < 0.01)were significantly higher in model group, while SOD, GSH-Px concentration reduced significantly (P < 0.01).
  
  
  CONCLUSIONS
  The in vitro cell model of steatosis and oxidative stress was successfully established. The activation of Nrf2/ARE pathway related factors maybe relevant to the overreaction of oxidative stress in HepG2 cells of steatosis.
authors.forename: Yu-pei
authors.initials: YP
authors.lastname: Zhang
authors.forename: Yi-lin
authors.initials: YL
authors.lastname: Kong
authors.forename: Qin-he
authors.initials: QH
authors.lastname: Yang
authors.forename: Ling
authors.initials: L
authors.lastname: Jin
authors.forename: Yin-ji
authors.initials: YJ
authors.lastname: Liang
authors.forename: Yi-fang
authors.initials: YF
authors.lastname: He
authors.forename: Yuan-jun
authors.initials: YJ
authors.lastname: Deng
authors.forename: Yuan-yuan
authors.initials: YY
authors.lastname: Li
authors.forename: Guan-long
authors.initials: GL
authors.lastname: Wang
authors.forename: Shao-bing
authors.initials: SB
authors.lastname: Cheng
mesh_terms: D006801:Humans; D061745:Antioxidant Response Elements; D003470:Culture Media; D005230:Fatty Acids, Nonesterified; D005234:Fatty Liver / Q000378:metabolism; D005979:Glutathione Peroxidase / Q000378:metabolism; D051547:Heme Oxygenase-1 / Q000378:metabolism; D056945:Hep G2 Cells; D008315:Malondialdehyde / Q000378:metabolism; D016660:NAD(P)H Dehydrogenase (Quinone) / Q000378:metabolism; D051267:NF-E2-Related Factor 2 / Q000378:metabolism; D009569:Nitric Oxide / Q000378:metabolism; D018384:Oxidative Stress; D017382:Reactive Oxygen Species / Q000378:metabolism; D013482:Superoxide Dismutase / Q000378:metabolism; D014280:Triglycerides / Q000378:metabolism
chemical_list: D003470:Culture Media; D005230:Fatty Acids, Nonesterified; D005979:Glutathione Peroxidase; D051547:Heme Oxygenase-1; C496541:HMOX1 protein, human; D008315:Malondialdehyde; D016660:NAD(P)H Dehydrogenase (Quinone); D051267:NF-E2-Related Factor 2; C495635:NFE2L2 protein, human; D009569:Nitric Oxide; C447479:NQO1 protein, human; D017382:Reactive Oxygen Species; D013482:Superoxide Dismutase; D014280:Triglycerides