【摘要】 目的:探讨长链非编码RNA（lncRNA）YAF2-AS1在肝星状细胞活化中的作用及可能的分子机制。方法:采用基因表达数据库（GEO）分析YAF2-AS1在肝纤维化组织和正常肝组织中的表达。用YAF2-AS1质粒转染人肝星状细胞LX-2细胞并设为YAF2-AS1组，以转染阴性对照质粒为对照组。采用CCK-8实验检测LX-2细胞增殖。采用流式细胞术检测LX-2细胞周期分布和活性氧（ROS）含量。采用lncRMap软件和双荧光素酶活性实验验证YAF2-AS1与miR-141-3p的靶向关系。采用实时荧光定量聚合酶链反应（qRT-PCR）检测miR-141-3p表达。采用Western blot检测蛋白磷酸酶及张力蛋白同源物（PTEN）/蛋白激酶B（AKT）信号通路中PTEN、p-AKT蛋白以及肝纤维化标志物[α平滑肌肌动蛋白（α-SMA）、Ⅰ型胶原蛋白（CollagenⅠ）、转化生长因子β受体Ⅱ（TGFβR2）]的表达。结果:与正常肝组织比较，肝纤维化组织YAF2-AS1表达水平降低（P<0.01）。与对照组比较，YAF2-AS1组LX-2细胞增殖活性被抑制（P<0.05）;G₀/G₁期LX-2细胞比例升高（P<0.01）,S期和G₂/M期LX-2细胞比例降低（均P<0.05）;LX-2细胞ROS含量升高（P<0.01）。miR-141-3p是YAF2-AS1的靶基因（P<0.01）。与对照组比较，YAF2-AS1组LX-2细胞miR-141-3p表达降低（P<0.01）;PTEN蛋白表达增加，p-AKT、α-SMA、CollagenⅠ、TGFβR2蛋白表达降低（均P<0.05）。结论:YAF2-AS1在肝纤维化组织中表达下调，过表达YAF2-AS1通过降低miR-141-3p表达抑制肝星状细胞的活化。更多还原

【Abstract】 Objective:To explore the role and possible molecular mechanism of long non-coding RNA（lncRNA） YAF2-AS1 in the activation of hepatic stellate cells. Methods: GEO database was used to analyze the expression of YAF2-AS1 in liver fibrosis tissue and normal liver tissue.Human hepatic stellate cell LX-2 cells were transfected with YAF2-AS1 plasmid and set as YAF2-AS1 group, and the negative control plasmid was used as the control group.CCK-8 assay was used to detect the proliferation of LX-2 cells.The cycle distribution and ROS content of LX-2 cells were detected by flow cytometry.LncRMap software and dual luciferase activity experiment were used to verify the targeting relationship between YAF2-AS1 and miR-141-3p.The expression of miR-141-3p was detected by qRT-PCR.Western blot was used to detect the expression of PTEN/AKT signaling pathway proteins（PTEN and p-AKT） and liver fibrosis markers（α-SMA, Collagen Ⅰ and TGFβR2）. Results: Compared with normal liver tissue, the expression level of YAF2-AS1 in liver fibrosis tissue was decreased（P<0.01）.Compared with control group, the proliferation activity of LX-2 cells in the YAF2-AS1 group was inhibited（P<0.05）; the proportion of LX-2 cells in G₀/G₁ phase was increased（P<0.01）,and the LX-2 cells in S and G₂/M phase were decreased（all P<0.05）;ROS content of LX-2 cells increased（P<0.01）.MiR-141-3p was the target gene of YAF2-AS1（P<0.01）.Compared with control group, the expression of miR-141-3p in LX-2 cells in YAF2-AS1 group was decreased（P<0.01）;the protein expression of PTEN was increased, and the protein expressions of p-AKT,α-SMA,CollagenⅠ and TGFβR2 were decreased（all P<0.05）. Conclusion:The expression of YAF2-AS1 is down-regulated in liver fibrosis tissue, and the overexpression of YAF2-AS1 inhibits the activation of hepatic stellate cells by reducing the expression of miR-141-3p.更多还原