摘要
目的探讨微小RNA-519d-3p(microRNA-519d-3p)对LX-2(人肝星状细胞株)活化及凋亡的影响。方法将miR-519d-3p分子模拟剂(miR-519d-3p mimics组)、抑制剂(miR-519d-3p inhibitor组)及其阴性对照物(mimics NC、inhibitor NC)转染LX-2细胞,利用实时定量PCR检测miRNA-519d-3p在转染后LX-2细胞中的相对表达量,利用实时定量PCR和Western blot检测转染后LX-2中活化标志物α平滑肌动蛋白(α-SMA)和Ⅰ型胶原蛋白(CollagenⅠ)的mRNA和蛋白的表达水平,分析miR-519d-3p对LX-2细胞活化的影响;用流式细胞术检测转染后LX-2的凋亡率,分析miR-519d-3p对LX-2细胞凋亡的影响。结果miR-519d-3p mimics组与其阴性对照组相比,α-SMA和Collagen I mRNA与蛋白表达水平均降低(P均<0.01);miR-519d-3p inhibitor组与其阴性对照组相比,α-SMA和Collagen I mRNA及蛋白表达水平均增高(P均<0.01);流式细胞术结果表明,miR-519d-3p mimics组可以促进细胞凋亡(P<0.01);miR-519d-3p inhibitor组可以抑制细胞凋亡,与阴性对照组相比差异均有统计学意义(P均<0.01)。结论miR-519d-3p具有抑制肝星状细胞活化,促进肝星状细胞凋亡,抑制肝纤维的作用。
Objective To investigate the effect of miR-519d-3p(microRNA-519d-3p)on activation and apoptosis of LX-2 cell line.Methods LX-2 cells(human hepatic stellate cell line)were transfected with miR-519d-3p molecular simulators(miR-519d-3p mimics)and inhibitors(miR-519d-3p inhibitor),and their negative controls(mimics NC,inhibitor NC)as negative controls.Real time quantitative PCR was used to detect the relative expression of miRNA-519d-3p in LX-2 cells.Real time PCR and Western blot were used to detect the mRNA and protein expression levels ofα-SMA and Collagen I in LX-2 cells after transfection,and the effect of mir-519d-3p on the activation of LX-2 cells was analyzed;the apoptosis rate of LX-2 cells after transfection was detected by flow cytometry,and the effect of mir-519d-3p on the apoptosis of LX-2 cells was analyzed.Results miR-519d-3p mimics could inhibitα-SMA and Collagen I mRNA expression level,compared with the control group,the difference was statistically significant(P all<0.01);miR-519d-3p inhibitor could promoteα-SMA and Collagen I mRNA expression level,the difference was statistically significant(P all<0.01);according to the results of Western blot,miR-519d-3p mimics could inhibitα-SMA and CollagenⅠprotein expression levels,compared with the control group,the difference was statistically significant(P<0.01);miR-519d-3p inhibitor could promoteα-SMA and CollagenⅠprotein expression levels,compared with the control group,the difference was statistically significant(P<0.01);Flow cytometry results showed that miR-519d-3p mimics could significantly promote apoptosis.On the contrary,miR-519d-3p inhibitor significantly inhibited apoptosis,with a statistically significant difference compared with the control group(P all<0.01).Conclusion miR-519d-3p can inhibit the activation of hepatic stellate cells,promote the apoptosis of hepatic stellate cells and inhibit the hepatic fiber.
作者
武彦虎
丁向春
王钊
冯薛烟
汪彭涛
海龙
WU Yanhu;DING Xiangchun;WANG Zhao;FENG Xueyan;WANG Pengtao;HAI Long(Ningxia Medical University,Yinchuan 750004,China;Department of Infectious Diseases,General Hospital of Ningxia Medical University,Yinchuan 750004,China;Laboratory of Pathogenic Microorganisms,General Hospital of Ningxia Medical University,Yinchuan 750004,China)
出处
《宁夏医科大学学报》
2021年第4期358-363,共6页
Journal of Ningxia Medical University
基金
国家自然科学基金(0101225)。
