miR-1247-3p激活TGF-β1信号介导肝星状细胞与肝纤维化的关系

MiR-1247-3p activates TGF-β1 signal to promote liver fibrosis via hepatic stellate cells

目的探讨miR-1247-3p对肝纤维化的影响,分析其对肝星状细胞增殖与活化的作用及分子机制。方法四氯化碳(CCl_(4))制备肝纤维化大鼠模型,并分离大鼠原代肝星状细胞。慢病毒转染处理细胞,分为空白对照组、miR-1247-3p mimic组和miR-1247-3p inhibitor组。检测miR-1247-3p、转化生长因子-β1(TGF-β1)、α-平滑肌肌动蛋白(α-SMA)水平及细胞增殖能力,并采用荧光素酶报告实验验证miR-1247-3p靶基因。结果与对照组比较,CCl_(4)腹腔注射后大鼠肝组织出现纤维化增生,结构破坏,炎症细胞浸润,且随时间延长肝纤维化程度加重(P<0.05);与对照组比较,模型组大鼠肝组织miR-1247-3p和TGF-β1表达水平均显著增加(P<0.05);与空白对照组比较,miR-1247-3p mimic组肝星状细胞增殖能力和肝星状细胞α-SMA蛋白表达水平显著增加,miR-1247-3p inhibitor组显著降低(P<0.05);与空白对照组比较,miR-1247-3p mimic组肝星状细胞TGF-β1表达水平显著升高,miR-1247-3p inhibitor组显著降低(P<0.05)。结论miR-1247-3p可通过靶向上调TGF-β1表达,促进肝星状细胞增殖和活化,从而参与肝纤维化进展。

Objective To investigate the effect of miR-1247-3p on hepatic fibrosis,and to analyze its effect on proliferation and activation of hepatic stellate cells and its molecular mechanism.Methods The rat model of hepatic fibrosis was established by carbon tetrachloride(CCl_(4)),and the primary hepatic stellate cells were isolated.The lentivirus transfected cells were divided into blank control group,miR-1247-3p mimic group and miR-1247-3p inhibitor group.The levels of miR-1247-3p,transforming growth factor-β1(TGF-β1),α-smooth muscle actin(α-SMA)and cell proliferation ability were detected,and the target gene of miR-1247-3p was verified by luciferase report experi-ment.Results Compared with the control group,hepatic fibrosis,structural destruction and infiltration of inflammatory cells appeared in rats after intraperitoneal injection of CCl_(4),and the degree of hepatic fibrosis increased with time(P<0.05).Compared with the control group,the expression levels of miR-1247-3p and TGF-β1 in the liver tissues of the model group were significantly increased(P<0.05).Compared with the blank control group,the proliferation ability andα-SMA protein expression level of hepatic stellate cells in miR-1247-3p mimic group were significantly increased,while those in miR-1247-3p inhibitor group were significantly reduced(P<0.05).TGF-β1 was the target gene of miR-1247-3p.compared with the blank control group,the expression level of TGF-β1 in hepatic stellate cells in miR-1247-3p mimic group was significantly increased,while that in miR-1247-3p inhibitor group was significantly reduced(P<0.05).Conclusion miR-1247-3p can promote the proliferation and activation of hepatic stellate cells by targeting up-regulating the expression of TGF-β1,thus participating in the progression of liver fibrosis.