miR-590-3p调控人肝星状细胞LX-2TGF-β/SMAD通路抗肝纤维化的研究

Effect of miR-590-3p on Anti-Liver Fibrosis by Regulating ECM and TGF-β/SMAD Pathway in Human LX-2 Cell Line

目的建立转化生长因子-β1(TGF-β1)诱导人肝星状细胞LX-2体外纤维化模型,观察miR-590-3p对肝纤维化影响。方法设计并合成miR-590-3p模拟物和miR-590-3p抑制剂,瞬时转染至LX-2细胞,再经TGF-β1诱导24 h后,荧光定量PCR检测胞内miR-590-3p表达,Western Blot检测胞内纤维化相关蛋白Smad 4及CollagenⅠ表达变化,并分析胞内miR-590-3p表达改变对细胞纤维化指标的影响。结果 miR-590-3p过表达下调Smad 4蛋白水平表达50%(P<0.05),抑制miR-590-3p功能上调Smad 4蛋白表达50%(P<0.05)。结论 miR-590-3p可能通过下调LX-2 Smad4的表达,影响LX-2活化,在肝纤维化进程中发挥重要作用。

OBJECTIVE To explore the establishment of a fibrosis model in vitro by inducing human LX-2 with transforming growth factor β1（ TGF-β1）,and to observe the effects of miR-590-3p on the expression of liver fibrosis markers. METHODS The miR-590-3p mimics and miR-590-3p inhibitor were designed,synthesized and transfacted into LX-2 cells. Transfacted cells were then induced by TGF-β1of 10 ng / m L for 24 h. The expression of miR-590-3p was detected by real-time PCR. The variations of expressions of fibrosis related protein Smad4 and Collagen Ⅰ were detected by the Western blotting to analyze the effects of the miR-590-3p expression on cell fibrosis indexes. RESULTS Compared to the control group,the expression of Smad4 protein significantly decreased 50% after LX-2 cells were transfacted with miR-590-3p mimics and increased 50% after LX-2 cells were transfacted with miR-590-3p inhibitor respectively（ P〈0. 05）. CONCLUSION The miR-590-3p may affect the activation of LX-2 cells through the depression of Smad 4 and then modulate liver fibrosis.