本文選題：心肌纖維化 + 成纖維細(xì)胞�。� 參考：《中國病理生理雜志》2017年10期

【摘要】：目的:研究微小RNA-199a-5p(miR-199a-5p)對心肌成纖維細(xì)胞中纖維化相關(guān)基因表達(dá)的調(diào)控作用及其可能作用的靶基因。方法:原代分離并體外培養(yǎng)成體C57BL/6小鼠心肌成纖維細(xì)胞;雙螢光素酶報(bào)告基因?qū)嶒?yàn)檢測miR-199a-5p與潛在靶基因沉默信息調(diào)節(jié)因子1(SIRT1)3’端非翻譯區(qū)(3’-UTR)的結(jié)合作用;實(shí)時(shí)熒光定量PCR(RT-q PCR)和Western blot法分別檢測SIRT1以及纖維化標(biāo)志物膠原蛋白(Col)1a1、Col3a1和α-平滑肌肌動蛋白(α-SMA)的mRNA和蛋白表達(dá)。結(jié)果:在血管緊張素Ⅱ(AngⅡ)誘導(dǎo)的小鼠心肌成纖維細(xì)胞中,Col1a1、Col3a1和α-SMA的表達(dá)增強(qiáng),miR-199a-5p表達(dá)上調(diào)。在心肌成纖維細(xì)胞中過表達(dá)miR-199a-5p可以增強(qiáng)Col1a1、Col3a1和α-SMA的表達(dá)。雙螢光素酶報(bào)告基因?qū)嶒?yàn)顯示miR-199a-5p與SIRT1 3’-UTR有結(jié)合作用。RT-q PCR和Western blot結(jié)果證實(shí)miR-199a-5p可在轉(zhuǎn)錄水平抑制SIRT1表達(dá)。過表達(dá)miR-199a-5p和沉默SIRT1均能一致性促進(jìn)心肌成纖維細(xì)胞中Col1a1、Col3a1和α-SMA的表達(dá)。抑制AngⅡ誘導(dǎo)的小鼠心肌成纖維細(xì)胞中NF-κB激活,可顯著降低miR-199a-5p表達(dá)。結(jié)論:SIRT1是miR-199a-5p的作用靶基因,并介導(dǎo)miR-199a-5p促進(jìn)纖維化標(biāo)志物Col1a1、Col3a1和α-SMA的表達(dá)。
[Abstract]:Aim: to study the regulatory effect of microRNA-199a-5pnmiR-199a-5p on the expression of fibrogenic genes in cardiac fibroblasts and the possible target genes. Methods: primary cultured adult C57BL / 6 mouse myocardial fibroblasts were isolated and cultured in vitro, and the binding of miR-199a-5p to potential target gene silencing information regulator 1SIRT1 3'untranslated region (3'UTRs) was detected by double luciferase reporter gene experiment. The mRNA and protein expressions of SIRT1, collagen marker Col1, Col3a1 and 偽 -smooth muscle actin (偽 -SMAs) were detected by real-time fluorescence quantitative PCR and Western blot, respectively. Results: the expression of Col1a1, Col3a1 and 偽 -SMA in myocardial fibroblasts induced by angiotensin 鈪，

本文編號：2044529