论文部分内容阅读
目的探讨微RNA-484(miR-484)对肝纤维化发生和发展过程中关键细胞肝星状细胞(HSC)的作用和影响。方法以前期芯片筛选结果为基础,通过细胞转染人为干预miR-484在HSC-T6细胞株中的表达。利用qPCR、蛋白质印迹法检测肝纤维化及凋亡相关指标的表达变化;以AnnexinⅤ-FITC/PI双染细胞后,用流式细胞仪检测miR-484对细胞凋亡的影响。结果与对照组相比,人为下调miR-484在HSC-T6细胞株中的表达后,其靶基因Fis1及促凋亡因子caspase-3在mRNA和蛋白水平的表达均升高(P<0.05),凋亡抑制因子Bcl-2表达下降(P<0.05);流式细胞仪检测到HSC-T6细胞凋亡率升高[(32.81±3.21)%vs(57.54±6.76)%,P<0.05];同时肝纤维化相关指标α-平滑肌肌动蛋白、Ⅰ型胶原的mRNA和蛋白水平均下调(P<0.05)。结论 miR-484通过靶向Fis1抑制HSC凋亡、促进其活化,从而促进肝纤维化的发生和发展。
Objective To investigate the effect of microRNA-484 (miR-484) on key hepatic stellate cells (HSC) during the development and progression of hepatic fibrosis. Methods Based on the previous screening results, miR-484 was transfected into HSC-T6 cells by human intervention. The changes of hepatic fibrosis and apoptosis related indexes were detected by qPCR and Western blotting. The apoptosis rate of miR-484 cells was detected by flow cytometry with AnnexinⅤ-FITC / PI double staining. Results Compared with the control group, miR-484 expression was down-regulated in HSC-T6 cells at both mRNA and protein levels (P <0.05) (32.81 ± 3.21)% vs (57.54 ± 6.76)%, P <0.05]. The apoptosis rate of HSC-T6 cells was significantly higher than that of HSC-T6 cells ; At the same time, the level of mRNA and protein of α-smooth muscle actin and collagen Ⅰ in liver fibrosis were down-regulated (P <0.05). Conclusion miR-484 can inhibit the HSC apoptosis by targeting Fis1 and promote its activation, thereby promoting the occurrence and development of hepatic fibrosis.