miR-143对HepG2细胞甘油三酯代谢的影响

Effects of miR-143 on lipid metabolism in Hep G2 cells

目的探讨miR-143对Hep G2细胞脂类代谢的调节作用。方法体外培养Hep G2细胞,待细胞密度达75%左右,在培养液中加入不同浓度游离脂肪酸(free fatty acid,FFA),油红O染色观察细胞内脂质积聚情况;采用miR-143mimics和Negative control(NC mimics)转染Hep G2细胞,实时荧光定量PCR检测miR-143的表达水平,评价miR-143转染效率;采用油红O染色观察miR-143过表达Hep G2细胞内脂质积聚情况;脂蛋白脂肪酶-甘油磷酸氧化酶-过氧化物酶-4-氨基安替比林和酚(GPO-PAP)法检测Hep G2细胞内甘油三酯(Triglyceride,TG)的含量。结果 0.25mmol/L FFA可导致Hep G2细胞内TG明显积聚,且呈时间依赖性;miR-143过表达可以显著抑制高FFA状态下Hep G2内TG的积聚。结论 miR-143可能参与了肥胖状态下肝脏细胞脂类代谢的调节。

Objective To investigate the role of miR-143 on lipid metabolism in Hep G2 cells. Methods Hep G2 cells were cultured in DMEM medium containing different concentrations of free fatty acid(FFA). miR-143 mimics and NC mimics were transfected into Hep G2 cells, which were evaluated using Quantitative real-time PCR(q RT-PCR) assays. The intracellular lipid droplets were observed by Oil red O staining, and triglycerides(TG) levels were detected using Lipoprotein lipase-glycerol phosphate oxidase-peroxidase 4-amino antipyrine and phenol(GPO-PAP) method. Results The contents of TG in Hep G2 cells treated with 0.25 m M FFA were significantly increased in a time-dependent manner. Overexpression of miR-143 significantly reduced the deposition of lipids in Hep G2 cells cultured with 0.25 m M FFA. Conclusion miR-143 may be involved in the dysregulation of liver lipid metabolism in obesity.