GATA-4基因促进5-氮杂胞苷诱导的骨髓间充质干细胞向心肌分化  

GATA-4 gene promote 5-azacytidine induced differentiation of bone marrow mesenchymal stem cells into cardiomyocytes

作　　者：张亚楠[1] 谢艳艳 尹明洁 张晓艺 陈炜[1] 张雷[1] ZHANG Ya-nan;XIE Yan-yan;YIN Ming-jie;ZHANG Xiao-yi;CHEN Wei;ZHANG Lei(Department of Histology and Embryology,the School of Basic Medical Sciences,Hebei Medical University,Shijiazhuang 050017,China;Class Ⅱ of Clinical Medicine,Grade 2015,the School of Basic Medical Sciences,Hebei Medical University,Shijiazhuang 050017,China)

机构地区：[1]河北医科大学基础医学院组织学与胚胎学教研室,河北石家庄050017 [2]河北医科大学基础医学院

出　　处：《河北医科大学学报》2019年第4期385-389,共5页Journal of Hebei Medical University

基　　金：河北省高等学校科学技术研究项目(QN2016159);河北省大学生创新性实验计划项目(USIP2017034)

摘　　要：目的探讨转染外源性基因GATA-4促进5-氮杂胞苷(5-azacytidine,5-aza)诱导的大鼠骨髓间充质干细胞(bone marrow mesenchymal stem cells,BMSCs)向心肌细胞分化的作用。方法全骨髓法分离大鼠BMSCs,按照1∶2.5比率用脂质体法将GATA-4(pVP22-GATA-4/myc-His)质粒瞬时转染第3代BMSCs,转染72 h后加入10μmol/L 5-aza诱导24 h,观察细胞形态变化。转染48 h后,免疫细胞化学检测GATA-4、myc的表达。于诱导后28 d免疫细胞化学及免疫印迹法检测GATA-4、心肌肌钙蛋白T(cardiac troponin T,cTnT)的表达情况,以鉴定细胞分化。结果转染72 h后,免疫细胞化学结果表明实验组可见GATA-4、myc表达。诱导后28 d应用免疫细胞化学及免疫印迹法检测结果显示,实验组GATA-4、cTnT表达显著高于对照组,差异有统计学意义(P<0.05)。结论外源表达GATA-4能够促进5-aza诱导的BMSCs向心肌分化。Objective To investigate the feasibility of GATA-4 gene promote 5-azacytidine(5-aza)induced differentiation of bone marrow mesenchymal stem cells(BMSCs)into cardiomyocytes.Methods Rat BMSCs were isolated from the whole bone marrow and cultured.Using the cationic liposome reagent,Lipofectamine 2000,the plasmid pVP22-GATA-4/myc-His was transfected into P3 BMSCs at a ratio of 1 ∶2.5.After 72 h,BMSCs was induced by 10 μmol/L 5-aza for 24 h.The whole experiments were observed under an inverted fluorescence microscope and expression of GATA-4 was examined with immunocytochemistry.Expression of cardiac troponin T(cTnT)and GATA-4 were determined with Western blotting and immunocytochemistry.Results After 48 h of transfection,GATA-4、myc were expressed in experimental group.After 28 days,immunocytochemistry and Western blotting results indicated that the expression of GATA-4 and cTnT in experimental group was significantly higher than those of control group(P<0.05).Conclusion GATA-4 gene may promote 5-aza induced differentiation of BMSCs into cardiomyocytes.

关 键 词：间质干细胞 骨髓 GATA4转录因子 细胞分化 

分 类 号：R394.2[医药卫生—医学遗传学]