【摘要】：目的探討微RNA-484(miR-484)對肝纖維化發(fā)生和發(fā)展過程中關(guān)鍵細胞肝星狀細胞(HSC)的作用和影響。方法以前期芯片篩選結(jié)果為基礎(chǔ),通過細胞轉(zhuǎn)染人為干預(yù)miR-484在HSC-T6細胞株中的表達。利用qPCR、蛋白質(zhì)印跡法檢測肝纖維化及凋亡相關(guān)指標的表達變化;以AnnexinⅤ-FITC/PI雙染細胞后,用流式細胞儀檢測miR-484對細胞凋亡的影響。結(jié)果與對照組相比,人為下調(diào)miR-484在HSC-T6細胞株中的表達后,其靶基因Fis1及促凋亡因子caspase-3在mRNA和蛋白水平的表達均升高(P0.05),凋亡抑制因子Bcl-2表達下降(P0.05);流式細胞儀檢測到HSC-T6細胞凋亡率升高[(32.81±3.21)%vs(57.54±6.76)%,P0.05];同時肝纖維化相關(guān)指標α-平滑肌肌動蛋白、Ⅰ型膠原的mRNA和蛋白水平均下調(diào)(P0.05)。結(jié)論 miR-484通過靶向Fis1抑制HSC凋亡、促進其活化,從而促進肝纖維化的發(fā)生和發(fā)展。
[Abstract]:Objective to investigate the effect and effect of microRNA-484 (miR-484) on hepatic stellate cell (HSC) (HSC) in the process of hepatic fibrosis. Methods based on the results of microarray screening, the expression of miR-484 in HSC-T6 cells was artificially interfered with by cell transfection. The expression of hepatic fibrosis and apoptosis-related indexes was detected by qPCR, Western blotting, and the effect of miR-484 on apoptosis was detected by flow cytometry after double staining of Annexin 鈪，

本文編號：2305949