河北医科大学学报

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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

  

  1. 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2015级临床医学二大班，河北 石家庄 050017
  • 出版日期:2019-04-25 发布日期:2019-04-22
  • 作者简介:张亚楠（1984-），女，河北石家庄人，河北医科大学基础医学院实验师，医学硕士，从事女性生殖系统研究。
  • 基金资助:
    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）

GATA4 gene promote 5azacytidine induced differentiation of bone marrow mesenchymal stem cells into cardiomyocytes#br#

  1. 1.Department of Histology and Embryology，the School of Basic Medical Sciences， Hebei Medical University，
    Shijiazhuang 050017， China； 2.Class Ⅱ of Clinical Medicine, Grade 2015， the School of
    Basic Medical Sciences， Hebei Medical University， Shijiazhuang 050017， China
  • Online:2019-04-25 Published:2019-04-22

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摘要： ［摘要］〓
〖HTH〗目的〖HTSS〗〖KG*2〗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。
〖HTH〗方法〖HTSS〗〖KG*2〗全骨髓法分离大鼠BMSCs，按照1〖DK〗∶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10 μmol/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。
〖HTH〗结果〖HTSS〗〖KG*2〗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）。
〖HTH〗结论〖HTSS〗〖KG*2〗外源表达GATA4能够促进5aza诱导的BMSCs向心肌分化。

关键词: 间质干细胞, 骨髓, GATA4转录因子, 细胞分化

Abstract: ［Abstract］〓Objective〖HTSS〗〓To investigate the feasibility of GATA4 gene promote 5azacytidine（5aza） induced differentiation of bone marrow mesenchymal stem cells(BMSCs) into cardiomyocytes.
〖HTH〗〖WTHZ〗Methods〖HTSS〗〓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〖DK〗∶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.
〖HTH〗〖WTHZ〗Results〖HTSS〗〓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）.
〖HTH〗〖WTHZ〗Conclusion〖HTSS〗〓GATA4 gene may promote 5aza induced differentiation of  BMSCs into cardiomyocytes.

Key words: mesenchymal stem cells, bone marrow, GATA4 transcription factor, cell differentiation

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