Effect of sirolimus on tumor necrosis factor α-induced lipid accumulation in HepG2 cells
10.13699/j.cnki.1001-6821.2017.06.013
- VernacularTitle:西罗莫司对肿瘤坏死因子α诱导的HepG2细胞脂质积聚的影响
- Author:
Bei-Bei LI
1
;
Ya-Xi CHEN
;
Xiong-Zhong RUAN
;
Lei ZHAO
Author Information
1. 重庆医科大学脂糖代谢性疾病重庆市重点实验室
- Keywords:
sirolimus;
non-alcoholic steatohepatitis;
mammalian target of rapamycin;
lipid accumulation
- From:
The Chinese Journal of Clinical Pharmacology
2017;33(6):526-529
- CountryChina
- Language:Chinese
-
Abstract:
Objective To investigate the effect of sirolimus on tumor necrosis factor alpha (TNF-o)-induced lipid accumulation in HepG2 cells.Methods Palmitate acid (PA)-loaded HepG2 cells were divided into three groups:Control group(0.2％ BSA + 0.16 mmol · mL-1 PA + 25 ng· mL-1 TNF-o),model group(0.2％ BSA +0.16 mmol · mL-1 PA +25 ng · mL-1TNF-α),experimental group(0.2％ BSA + 0.16 mmol · mL-1 PA +25 ng · mL-1 TNF-α + 10 ng · mL-1 sirolimus).After 24 h treatment,lipid accumulation in the HepG2 cells was visualized using oil red O staining.The phosphorylation of mammalian target of rapamycin (mTOR)and its downstream translation regulator including P70 ribosomal protein S6 kinase (P70S6K) in HepG2 cells were detected by Western blotting.The mRNA expression of sterol regulatory element binding protein 1 (SREBP1),acetyl CoA carboxylase(ACC),fatty acid synthetase(FAS) were detected by real-time PCR.Results TNF-o-induced lipid accumulation was significantly inhibited by sirolimus in HepG2 cells.The TNF-α-induced phosphorylation of mTOR and its downstream translation regulator P70S6K were also reduced by sirolimus:the relative levels of p-mTOR protein in control group,model group and experimental group were 1.00 ± 0.20,3.11 ± 0.60,2.38 ± 0.50,respectively;the relative levels of p-P70S6K protein in that three groups were 1.00 ±0.30,3.67 ±0.60,1.62 ±0.50,respectively.The differences between model group and control group or experimental group and model group were statistically significant (all P ＜ 0.05).Additionally,the mRNA expression of SREBP1,ACC and FAS were downregulated by sirolimus:the mRNA levels of FAS in control group,model group andexperimental group were 1.04 ± 0.32,2.85 ±0.90,1.68 ± 0.38,respectively;the mRNA levels of ACC in that three groups were 1.04 ± 0.30,3.23-±1.33,2.07 ± 0.52,respectively;the mRNA levels of SREBP1 in that three groups were 1.01 ± 0.16,3.85 ± 1.30,2.82 ±0.57,respectively.The differences between model group and control group or experimental group and model group were statistically significant (all P ＜ 0.05).Conclusion Sirolimus ameolirates TNF-α-induced lipid accumulation in HepG2 cells through inhibiting the mTOR signaling pathway.
