河北医科大学学报

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过表达GATA4的骨髓间充质干细胞向心肌细胞分化的研究

  

  1. 1.河北医科大学基础医学院组织胚胎学教研室，河北 石家庄 050017；2.河北医科大学基础医学院病原学教研室，河北 石家庄 050017
  • 出版日期:2019-08-25 发布日期:2019-08-14
  • 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，医学博士研究生，从事组织胚胎学研究。
  • 基金资助:
    河北省高等学校科学研究计划（QN2016159）；河北省大学生创新性实验计划项目（USIP2017034）

Study on differentiation from bone marrow mesenchymal stem cells overexpressing GATA4 into cardiomyocytes

  1. 1.Department of Histology and Embryology， the School of Basic Medicine Sciences， Hebei Medical
    University， Shijiazhuang 050017， China； 2.Department of  Pathogenic and Microorganism， the
    School of Basic Medicine Sciences， Hebei Medical University， Shijiazhuang 050017， China
  • Online:2019-08-25 Published:2019-08-14

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摘要： ［摘要］〓
〖HTH〗目的〖HTSS〗〖KG*2〗探讨过表达GATA4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，经多次传代纯化后，借助细胞穿透肽VP22的载体作用，采用脂质体转染法将GATA4(pVP22GATA4/mycHis)基因瞬时转染SD大鼠BMSCs，于转染后第3天加入适宜浓度的5氮杂胞苷（5azacytidine，5aza）诱导1 d，同时观察细胞形态变化。转染2 d后，采用免疫印迹法检测GATA4、myc的表达。于诱导后21 d采用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，以鉴定细胞分化。
〖HTH〗结果〖HTSS〗〖KG*2〗 转染2 d后，免疫印迹法检测结果显示实验组有GATA4、myc表达。诱导后21 d，免疫印迹法、免疫细胞化学法检测结果显示实验组GATA4、cTnT表达水平显著高于空质粒转染组和空白对照组（P＜005）。
〖HTH〗结论〖HTSS〗〖KG*2〗过表达GATA4基因能够促进经5aza诱导的BMSCs向心肌细胞分化。

关键词: 间质干细胞, 骨髓, 肌细胞, 心脏, GATA4

Abstract: ［Abstract］〓Objective〖HTSS〗〓To investigate the role of Bone Marrow Mesenchymal Stem Cells(BMSCs) that overexpression GATA4 gene in differentiating into cardiomyocytes.
〖HTH〗〖WTHZ〗Methods〖HTSS〗〓Rat BMSCs were isolated from the whole bone marrow and cultured.After multiple passages and purification，the GATA4 (pVP22GATA4/mycHis) gene was transiently transfected into SD rat BMSCs by liposome transfection with the help of the cellpenetrating peptide VP22. After 3 days, BMSCs was induced by a suitable concentration of 5aza for 1 day. The expression of GATA4 and myc were examined by Western blotting. Expression of cardiac troponin T(cTnT) and GATA4 were determined with immunocytochemistry and Western blotting after 21 days.
〖HTH〗〖WTHZ〗Results〖HTSS〗〓GATA4 and myc were expressed in experimental group after 2 days of transfection. After 21 days, Western blotting and immunocytochemistry results indicated that the expression of GATA4 and cTnT in experimental group was significantly higher than those other groups.
〖HTH〗〖WTHZ〗Conclusion〖HTSS〗〓Overexpression GATA4 gene could promote BMSCs differentiate into cardiomyocytes during 5aza induction.

Key words: mesenchymal stem cells, bone marrow, myocytes, cardiac, GATA4

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