【摘要】 目的探讨过表达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。方法全骨髓法分离大鼠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。结果转染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)。结论过表达GATA-4基因能够促进经5-aza诱导的BMSCs向心肌细胞分化。更多还原

【Abstract】 Objective To investigate the role of Bone Marrow Mesenchymal Stem Cells(BMSCs) that overexpression GATA-4 gene in differentiating into cardiomyocytes. Methods 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. Results 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. Conclusion Overexpression GATA-4 gene could promote BMSCs differentiate into cardiomyocytes during 5-aza induction.更多还原