本文選題：LncRNA + GAS��； 參考：《安徽醫(yī)科大學學報》2017年03期

【摘要】：目的應用長鏈非編碼RNA GAS5(LncRNA GAS5)過表達質(zhì)粒(pEGFP-C1-GAS5)轉(zhuǎn)染SD大鼠心肌成纖維細胞,觀察其對心肌成纖維細胞活化增殖的影響。方法提取SD大鼠心肌成纖維細胞,轉(zhuǎn)化生長因子β1(TGF-β1)刺激其增殖,分為pEGFP-C1-GAS5組、陰性對照組和空白對照組,應用LncRNA GAS5基因的過表達質(zhì)粒(pEGFP-C1-GAS5)通過脂質(zhì)體瞬時轉(zhuǎn)染細胞,48 h后收獲細胞。采用實時定量聚合酶鏈反應(qRT-PCR)檢測GAS5、Ⅰ型膠原前膠原(Col1A1)和α-平滑肌肌動蛋白(α-SMA)的mRNA表達,Western blot分析Col1A1和α-SMA蛋白表達變化,MTT法檢測細胞增殖活力。結果 TGF-β1刺激心肌成纖維細胞后,qRT-PCR結果顯示LncRNA GAS5的表達量較正常組下降(P0.05);與陰性對照組和空白對照組比較,轉(zhuǎn)染了pEG-FP-C1-GAS5的實驗組,qRT-PCR結果顯示實驗組的GAS5表達均明顯升高(P0.05),同時α-SMA和Col1A1 mRNA表達顯著降低(P0.05);Western blot結果顯示轉(zhuǎn)染pEG-FP-C1-GAS5的實驗組α-SMA和Col1A1蛋白表達量均明顯降低(P0.05);MTT法檢測結果表明在實驗組心肌成纖維細胞的增殖活力低于陰性對照組和空白對照組(P0.05)。結論過表達LncRNA GAS5可顯著降低心肌成纖維細胞的增殖活力,提示GAS5有抑制心肌成纖維細胞的活化增殖的作用,為以后進一步研究提供依據(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：1977310