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[Expression and significance of Nrf2/ARE pathway ralated factors in the HepG2 cell model of steatosis].
- Source :
-
Zhongguo ying yong sheng li xue za zhi = Zhongguo yingyong shenglixue zazhi = Chinese journal of applied physiology [Zhongguo Ying Yong Sheng Li Xue Za Zhi] 2016 Jan; Vol. 32 (1), pp. 13-7. - Publication Year :
- 2016
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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.<br />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<br />Nad(p)h: quinone oxidoreductase-1(NQO1) were analyzed by Western blot.<br />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).<br />Conclusion: 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.
- Subjects :
- Humans
Culture Media
Fatty Acids, Nonesterified
Glutathione Peroxidase metabolism
Heme Oxygenase-1 metabolism
Hep G2 Cells
Malondialdehyde metabolism
NAD(P)H Dehydrogenase (Quinone) metabolism
Nitric Oxide metabolism
Reactive Oxygen Species metabolism
Superoxide Dismutase metabolism
Triglycerides metabolism
Antioxidant Response Elements
Fatty Liver metabolism
NF-E2-Related Factor 2 metabolism
Oxidative Stress
Subjects
Details
- Language :
- Chinese
- ISSN :
- 1000-6834
- Volume :
- 32
- Issue :
- 1
- Database :
- MEDLINE
- Journal :
- Zhongguo ying yong sheng li xue za zhi = Zhongguo yingyong shenglixue zazhi = Chinese journal of applied physiology
- Publication Type :
- Academic Journal
- Accession number :
- 27255033