本文選題：LncRNA + GAS　； 參考：《安徽醫(yī)科大學(xué)學(xué)報(bào)》2017年03期

【摘要】：目的應(yīng)用長(zhǎng)鏈非編碼RNA GAS5(LncRNA GAS5)過(guò)表達(dá)質(zhì)粒(pEGFP-C1-GAS5)轉(zhuǎn)染SD大鼠心肌成纖維細(xì)胞,觀(guān)察其對(duì)心肌成纖維細(xì)胞活化增殖的影響。方法提取SD大鼠心肌成纖維細(xì)胞,轉(zhuǎn)化生長(zhǎng)因子β1(TGF-β1)刺激其增殖,分為pEGFP-C1-GAS5組、陰性對(duì)照組和空白對(duì)照組,應(yīng)用LncRNA GAS5基因的過(guò)表達(dá)質(zhì)粒(pEGFP-C1-GAS5)通過(guò)脂質(zhì)體瞬時(shí)轉(zhuǎn)染細(xì)胞,48 h后收獲細(xì)胞。采用實(shí)時(shí)定量聚合酶鏈反應(yīng)(qRT-PCR)檢測(cè)GAS5、Ⅰ型膠原前膠原(Col1A1)和α-平滑肌肌動(dòng)蛋白(α-SMA)的mRNA表達(dá),Western blot分析Col1A1和α-SMA蛋白表達(dá)變化,MTT法檢測(cè)細(xì)胞增殖活力。結(jié)果 TGF-β1刺激心肌成纖維細(xì)胞后,qRT-PCR結(jié)果顯示LncRNA GAS5的表達(dá)量較正常組下降(P0.05);與陰性對(duì)照組和空白對(duì)照組比較,轉(zhuǎn)染了pEG-FP-C1-GAS5的實(shí)驗(yàn)組,qRT-PCR結(jié)果顯示實(shí)驗(yàn)組的GAS5表達(dá)均明顯升高(P0.05),同時(shí)α-SMA和Col1A1 mRNA表達(dá)顯著降低(P0.05);Western blot結(jié)果顯示轉(zhuǎn)染pEG-FP-C1-GAS5的實(shí)驗(yàn)組α-SMA和Col1A1蛋白表達(dá)量均明顯降低(P0.05);MTT法檢測(cè)結(jié)果表明在實(shí)驗(yàn)組心肌成纖維細(xì)胞的增殖活力低于陰性對(duì)照組和空白對(duì)照組(P0.05)。結(jié)論過(guò)表達(dá)LncRNA GAS5可顯著降低心肌成纖維細(xì)胞的增殖活力,提示GAS5有抑制心肌成纖維細(xì)胞的活化增殖的作用,為以后進(jìn)一步研究提供依據(jù)。
[Abstract]:Objective to investigate the effect of pEGFP-C1-GAS5 (long chain non-coding RNA GAS5(LncRNA GAS5) overexpression plasmid (pEGFP-C1-GAS5) on the activation and proliferation of myocardial fibroblasts in SD rats. Methods Sprague-Dawley rat myocardial fibroblasts were extracted and stimulated by TGF- 尾 1 (TGF- 尾 1). They were divided into pEGFP-C1-GAS5 group, negative control group and blank control group. The LncRNA GAS5 gene overexpression plasmid pEGFP-C1-GAS5 was used to transfect the cells through liposome for 48 h. The expression of GAS5, procollagen 鈪，

本文編號(hào)：1977310