微小RNA-214对肝纤维化的影响

Effects of microRNA-214 on liver fibrosis

目的探讨miRNA-214对肝纤维化的影响及其可能的机制。方法通过腹腔注射CCl4构建小鼠肝纤维化模型,运用逆转录-聚合酶链反应(reversetranscriotion-polymerasechainreaction,RT-PCR)法检测miR-214的表达情况。通过脂质体转染方式调控LX-2细胞miR-214的表达水平,采用CCK-8法、RT-PCR及划痕实验检测转染miR-214模拟物和miR-214抑制物后LX-2细胞增殖、活化及迁移能力的变化。通过蛋白质印迹法对数据库Targetscan预测的miR-214靶基因进行验证。结果模型组小鼠肝脏miR-214表达水平显著高于对照组(t=1.928,P=0.040)。LX-2细胞转染miR-214模拟物后,miR-214表达上调(t=3.872,P=0.001);LX-2细胞α-SMA、胶原ⅠαmRNA的表达水平上调(t=1.952,P=0.039;t=2.926,P=0.006),LX-2细胞增殖和迁移能力增强,数据库Targetscan预测的靶基因PTEN蛋白表达量下调。结论 miR-214参与HSC的活化、增殖及迁移,可促进肝纤维化的形成,miR-214可能是通过抑制PTEN的表达来影响HSC的生物学功能。

Objective To investigate the effects of miR-24 on liver fibrosis in vivo and in vitro. MethodsThe model of liver fibrosis was established by intraperitoneal injection of CCl4 into C57BL/6 mice and realtimePCR was used to determinate the expression of miR-214. The expression of miR-214 in LX-2 cellswas regulated by liposome transfection, and the proliferation, activation and migration of LX-2 cells aftertransfection of miR-214 mimics and miR-214 inhibitor were investigated by CCK-8, reverse transcriotionpolymerasechain reaction （RT-PCR） and scratch test. The potential target gene of miR-214 was predicted inthe Targetscan database, and was verified by Western-blot. Results The expression of miR-214 in the liver ofthe model group was significantly higher than that of the control group （t = 1.928,P = 0.040）. Compared withthe NC-mimics, the miR-214 mimics can promote the LX-2 cell proliferation and migration （t = 3.872, P =0.001）, increase the mRNA expression of α-SMA （t = 1.952, P = 0.039） and collagen Ⅰα （t = 2.926, P = 0.006）.The miR-214 mimics could also decrease the protein expression of PTEN which were predicted by Targetscandatabase. Conclusions MiR-214 is involved in the activation, proliferation and migration of HSC and promotesthe formation of liver fibrosis. It is possible that the biological function of HSC is affected by inhibiting theexpression of PTEN.