miR-369-5p对SD大鼠心肌成纤维细胞活化增殖的影响  

Effect of miR-369-5p on cardiac fibroblast activationand proliferation in SD rats

作　　者：代晨[1,2] 陶辉[1,2] 石开虎[1,2] 徐盛松[1,2] DAI Chen;TAO Hui;SHI Kai-hu;XU Sheng-song(Dept of Cardiothoracic Surgery,the Second Hospital of Anhui Medical University,Hefei 230601,China;Cardiovascular Research Center,Anhui Medical University,Hefei 230601,China)

机构地区：[1]安徽医科大学第二附属医院心胸外科,安徽合肥230601 [2]安徽医科大学心血管病研究中心,安徽合肥230601

出　　处：《中国药理学通报》2018年第12期1693-1697,共5页Chinese Pharmacological Bulletin

基　　金：国家自然科学基金资助项目(No 81570295);安徽省科技攻关资助项目(No 1501041148);安徽省高校自然科学研究项目(No KJ2017A168);安徽省自然科学基金资助项目(No 1808085MH231)

摘　　要：目的探讨miR-369-5p对大鼠心肌成纤维细胞(cardiac fibroblasts,CFs)活化增殖的影响。方法差速贴壁离心法提取CFs,并于倒置相差显微镜下进行形态鉴定。分别向各组细胞瞬时转染miR-369-5p抑制物、miR-369-5p模拟物、阴性对照组(NC)24～48 h后,应用实时定量PCR检测miR-369-5p、α-平滑肌肌动蛋白(α-SMA)和I型胶原前胶原A1(Col1A1)的蛋白表达;CCK-8法检测细胞增殖活性;Western blot测定α-SMA和Col1A1蛋白表达。结果 miR-369-5p在模拟物组CFs中表达上调;而在抑制物组中的表达下降。Western blot结果显示,α-SMA和Col1A1蛋白在模拟物组中的表达明显降低,在抑制物组中的表达明显增加。CFs瞬时转染miR-369-5p模拟物后培养48 h,CFs增殖活力明显下降,而抑制物组的CFs活力明显增强。结论 miR-369-5p表达下调能够促进CFs的活化增殖,提示miR-369-5p是心肌纤维化的可能作用靶点。Aim To investigate the effect of miR-369-5p on activation and proliferation of rat cardiac fibroblasts （CFs）. Methods Myocardial fibroblasts were extracted by differential adherent centrifugation and identified by inverted phase contrast microscopy. The miR-369-5p inhibitors, miR-369-5p mimics and liposomal blank control groups were transiently transfected into each group of cells for 24 h to 48 h. miR-369-5p, α-smooth muscle actin （α-SMA） and type I collagen procollagen A1 （Col1A1） gene expression were detected by real-time quantitative PCR; cell proliferation activity was assessed by CCK-8 assay; α-SMA and Col1A1 protein expression was detected by Western blot. Results miR-369-5p was up-regulated in CFs in mimics group, but decreased in inhibitors group. Western blot showed that the expression of α-SMA and Col1A1 protein in the mimics group significantly decreased, while the expression in inhibitors group significantly increased. After 48 h of transient transfection of miR-369-5p mimics by CFs, the proliferation activity of CFs significantly decreased, while the activity of CFs in the inhibitors group significantly increased. Conclusions miR-369-5p mimics can significantly inhibit the proliferation of CFs, while miR-369-5p inhibitors can promote the proliferation of CFs, suggesting that miR-369-5p may be a target of myocardial fibrosis.

关 键 词：心肌成纤维细胞 心肌纤维化 miR-369-5p 增殖 Α-平滑肌肌动蛋白 COL1A1 

分 类 号：R-332[医药卫生] R322.11