Expression and significance of Nrf2/ARE pathway ralated factors in the HepG2 cell model of steatosis.
- Author:
Yu-pei ZHANG
;
Yi-lin KONG
;
Qin-he YANG
;
Ling JIN
;
Yin-ji LIANG
;
Yi-fang HE
;
Yuan-jun DENG
;
Yuan-yuan LI
;
Guan-long WANG
;
Shao-bing CHENG
- Publication Type:Journal Article
- MeSH: Antioxidant Response Elements; Culture Media; Fatty Acids, Nonesterified; Fatty Liver; metabolism; GA-Binding Protein Transcription Factor; Glutathione Peroxidase; metabolism; Heme Oxygenase-1; metabolism; Hep G2 Cells; Humans; Malondialdehyde; metabolism; NAD(P)H Dehydrogenase (Quinone); metabolism; NF-E2-Related Factor 2; metabolism; Nitric Oxide; metabolism; Oxidative Stress; Reactive Oxygen Species; metabolism; Superoxide Dismutase; metabolism; Triglycerides; metabolism
- From: Chinese Journal of Applied Physiology 2016;32(1):13-17
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo 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.
METHODSHepG2 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
NAD(P)Hquinone oxidoreductase-1(NQO1) were analyzed by Western blot.
RESULTSCompared 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).
CONCLUSIONThe 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.