LncRNA-ZFAS1对心肌纤维化调控作用研究

目的探究lncRNA-ZFAS1对心肌纤维化的调控作用。方法采用结扎小鼠心脏冠状动脉左前降支的方式建立心肌缺血诱发的心肌纤维化模型;利用CRISPR/Cas9技术构建心脏特异性过表达lncRNA-ZFAS1的转基因小鼠;利用Masson染色检测小鼠心脏组织心肌纤维化情况;利用血管紧张素Ⅱ(AngⅡ)诱导原代培养的小鼠心肌成纤维细胞,建立离体心肌纤维化模型,并转染siRNA(siZFAS1)用于敲减lncRNA-ZFAS1;利用MTT法检测心肌成纤维细胞增殖情况;利用实时荧光定量PCR(qRT-PCR)实验检测lncRNA-ZFAS1、转化生长因子β1(TGFβ1)、Ⅰ型胶原(Col1a1)和Ⅲ型胶原(Col3a1)的表达情况。利用Western blot实验检测α-平滑肌肌动蛋白(alpha-smooth muscle actin,α-SMΑ)的表达情况。结果在心肌纤维化模型小鼠的心脏组织中,lncRNA-ZFAS1的表达显著升高,AngⅡ诱导的心肌成纤维细胞中敲减lncRNA-ZFAS1后,Col3a1和TGFβ1的表达显著降低;心脏特异性过表达lncRNA-ZFAS1的转基因小鼠心脏组织中出现胶原沉积,并且α-SMA、Col1a1和Col3a1的表达显著升高。结论lncRNA-ZFAS1在心脏组织中的高表达会诱导心肌纤维化,敲减lncRNA-ZFAS1可以缓解心肌纤维化。

Research on regulatory effect of lncRNA-ZFAS1 on myocardial fibrosis

Aim To explore the regulatory effect of lncRNA-ZFAS1 on myocardial fibrosis.Methods The mouse model of myocardial fibrosis was established by ligating the left anterior descending coronary artery of heart;the lncRNA-ZFAS1 cardiac-specific transgenic mice were constructed by CRISPR/Cas9 technology.Masson staining was used to detect the fibrosis of heart tissues;angiotensinⅡ(AngⅡ)was added to cardiac fibroblasts to establish an in vitro myocardial fibrosis model,and siRNA was transfected to knock down lncRNA-ZFAS1;MTT method was used to detect the proliferation of cardiac fibroblasts;real-time quantitative PCR(qRT-PCR)was used to detect the expression of lncRNA-ZFAS1,transforming growth factorβ1(TGFβ1),type I collagen(Col1a1)and typeⅢcollagen(Col3a1).Western blot was used to detectɑ-SMA expression.Results In cardiac tissues of myocardial fibrosis model mice,the expression of lncRNA-ZFAS1 significantly increased.Knocking down of lncRNA-ZFAS1significantly reduced the expression of Col3a1 and TGFβ1 in AngⅡtreated cardiac fibroblasts.Collagen deposition appeared in cardiac tissues of lncRNA-ZFAS1 transgenic mice,and the expression ofα-SMA,Col1a1 and Col3a1 markedly increased.Conclusions The high expression of lncRNA-ZFAS1 in heart tissues could induce myocardial fibrosis,and knocking down lncRNA-ZFAS1 could alleviate myocardial fibrosis.