Objective: To investigate the role and probable mechanism of miRNA-181a in nonalcoholic fatty liver disease. Methods: HepG2 cells were treated with palmitic acid to construct a nonalcoholic fatty liver disease cell model, and the expression of miR-181a and lipidosis in the cells were measured. Transforming growth factor-β (TGF-β) was used to examine the effect of miR-181a expression in HepG2 cells. The miR-181a, lipidosis, reduced glutathione and reactive oxygen species (ROS) were determined by controlling and regulating the miR-183 expression levels after transfection with miR-181 mimics and inhibitors in HepG2 cells. The miR-181a target genes were predicted by bioinformatics analysis, and verified by real-time fluorescent quantitative PCR and western blotting. The independent sample t-test was used for the comparison between the two independent samples, and the comparison between multiple groups were accorded with the normal distribution, homogeneity of variance, and one-way analysis of variance. Results: Lipidosis was significantly increased after palmitic acid treatment in HepG2 cells, and the expression level of miR-181a was significantly increased than control group. After HepG2 cells were transfected with miR-181a inhibitors, the expression of miR-181a, triglycerides and reactive oxygen species were down-regulated, and reduced glutathione, predicting the mRNA and protein expression of target gene silencing information regulator 2 related enzyme 1 were up-regulated. However, the results were contrary to the above changes after transfection with miR-181a mimics. Conclusion: miR-181a participates in lipidosis and promotes lipid peroxidation in nonalcoholic fatty liver disease. miR-181a may affect the pathogenesis and progression of nonalcoholic fatty liver disease by inhibiting the expression of silencing information regulator 2 related enzyme 1.
目的： 探讨miRNA-181a（miR-181a）对非酒精性脂肪性肝病的影响及其可能的机制。 方法： 使用棕榈酸处理HepG2细胞构建非酒精性脂肪性肝病细胞模型，检测细胞内miR-181a的表达及脂质沉积。用转化生长因子-β处理细胞检测其对miR-181a表达的影响。通过调控HepG2细胞miR-181a的表达水平，分别检测转染miR-181a模拟物和抑制物后HepG2细胞内miR-181a表达、脂质沉积和还原型谷胱甘肽和活性氧水平。通过实时荧光定量PCR和蛋白质印迹法对生物信息学预测的miR-181a靶基因进行验证。两组独立样本间比较采用独立样本t检验，多组间比较资料特征符合正态分布且方差齐性，采用单因素方差分析。 结果： 棕榈酸处理后HepG2中脂质沉积明显增多，miR-181a表达水平显著高于对照组。HepG2细胞转染miR-181a抑制物后，miR-181a表达下调，甘油三酯和活性氧下降，还原型谷胱甘肽升高，预测靶基因沉默信息调节因子2相关酶1的mRNA和蛋白表达量上调；而转染miR-181a模拟物后，结果与上述改变相反。 结论： miR-181a参与非酒精性脂肪性肝病的脂质沉积并促进脂质过氧化，miR-181a可能是通过抑制沉默信息调节因子2相关酶1的表达来影响非酒精性脂肪性肝病的发病机制及进展。.
Keywords: Non-alcoholic fatty liver disease; Sirtuin1; Transforming growth factor; miR-181a.