LncRNA-Braveheart促进骨髓间充质干细胞在体外向心肌样细胞分化
LncRNA-Braveheart promotes the differentiation of bone marrow mesenchymal stem cells in vitro into cardiomyocyte-like cells
摘要
Abstract
BACKGROUND:Our previous work demonstrated that bone marrow mesenchymal stem cells (BMSCs) transplantation could improve cardiac function in rats with myocardial infarction.However,the overall efficacy was unsatisfactory,and there was a low efficiency of BMSCs differentiating into cardiomyocytes in the local infarct myocardium.OBJECTIVE:To transfect long non-coding RNA-Braveheart (IncRNA-Bvht) into BMSCs in order to observe whether it could promote cardiomyocyte differentiation of BMSCs in vitro.METHODS:pLVX-IRES-ZsGreen1-IncRNA-Bvht vector was constructed and applied to transfect IncRNA-Bvht into bMSCs,and then,the transfection efficiency was detected.BMSCs were obtained from C57BL/6 mice and cultured ir vitro.Passage 3 cells were divided into three groups:BMSCs group,null vector group and IncRNA-Bvht group.All cells in the three groups were cultured in the normal condition for 48 hours and cardiomyocytes differentiation was induced by 5-azacytidine for another 24 hours followed by 2-week culture under normal conditions.Cardiomyocyte differentiation of BMSCs was observed under fluorescence microscopy and expression of cardiac specific cell markers including troponin T and myosin were examined using immunofluorescent staining,western blot assay,and qRT-PCR.RESULTS AND CONCLUSION:Cell morphological changes could be observed in all the groups 2 weeks after the induction.Interconnected cells arranged consistently in all the three groups.Immunofiuorescent staining results showed that the expression of troponin T and myosin was notably positive,and the proportion of troponin T positive cells was significantly increased.qRT-PCR and western blot assay results indicated that there were significantly increased levels of troponin T and myosin in the IncRNA-Bvht group as compared with the BMSCs and null vector groups (P ＜ 0.01),suggesting that IncRNA-Bvht could efficiently promote cardiomyocyte differentiation of BMSCs in vitro.关键词
干细胞/骨髓间充质干细胞/长链非编码RNA-Braveheart/心肌样细胞/心肌梗死/国家自然科学基金分类
医药卫生引用本文复制引用
侯婧瑛,王彤,龙会宝,周长青,吴浩,郭天柱,钟婷婷,伍权华,汪蕾,郑韶欣..LncRNA-Braveheart促进骨髓间充质干细胞在体外向心肌样细胞分化[J].中国组织工程研究,2017,21(29):4593-4599,7.基金项目
国家自然科学基金(81070125)“抗凋亡与促血管生成miRNA-378干预MSCs治疗心梗后心衰的机制研究” （81070125）
国家自然科学基金(81270213)“ANGⅡ/AT1/SMAD/C×43通路在心肌干细胞提高心梗大鼠心电生理学稳定性和室颤阈值的作用机制研究” （81270213）
国家自然科学基金(81670306)“心肌干细胞经IGF-1/STAT/miR-155通路下调4T1R改善心肌梗死后心电生理稳定性机理研究” （81670306）
广东省科技计划项目 (2014A020211002)“LncRNA-Bvht/MESP1/N-cadherin通路调控MSCs向心肌细胞定向分化的机制研究” （2014A020211002）
广东省科技计划项目(2010B031600032)“抗凋亡与促血管生成niRNA-378干预MSCs治疗心梗后心衰的机制研究” （2010B031600032）
高校基本科研业务费中山大学青年教师重点培育项目(13ykzd16)“PPAR-γ/TGF-β1/Smad/CX43通路在PPAR-γ干预MSCs治疗心梗后心衰的疗效及机制研究” （13ykzd16）
广东省医学科研基金(A2016264)“心肌干细胞经由HIF-1α/Apelin/APJ/ACE2通路下调ANGII改善心肌梗死大鼠心电生理学稳定性的机制研究” （A2016264）
the National Natural Science Foundation of China,No.81070125,81270213,81670306 （）
the Science and Technology Plan Projects of Guangdong Province,No.2014A020211002,2010B031600032 （）
Fundamental Research Fund for Universities-Young Teacher Training Project of Sun Yat-Sen University,No.13ykzd16 （）
the Medical Research Fund of Guangdong Province,No.A2016264 （）
