【摘要】 目的观察骨髓间充质干细胞(MSCs)向心肌诱导分化过程中,心肌转录因子GATA-4和Nkx2.5基因表达的变化规律,探讨其分子生物学机理。方法体外培养大鼠骨髓间充质干细胞,取第3代以10μmol/L 5-氮杂胞苷(5-aza)诱导,分别于第1、2、3、4周收集细胞,免疫细胞化学法和Western blot法检测肌钙蛋白I(troponinI,TnI)表达,RT-PCR法分析GATA-4和Nkx2.5基因的表达。结果诱导后细胞停止生长,并倾向集落化,诱导第2周开始出现TnI阳性细胞,第1周开始即有GATA-4和Nkx2.5基因表达,逐渐增强,第3周达到高峰,持续到第4周。结论GATA-4和Nkx2.5基因表达在骨髓间充质干细胞向心肌样细胞诱导过程中逐渐增强,可能参与调控心肌细胞的分化过程。更多还原

【Abstract】 Objective To investigate the expression of the transcriptive factors,GATA-4 and Nkx2.5 gene,in the course of induction of mesenchymal stem cells(MSCs) into cardiomyocytes,and analyze it’s molecular biological mechanism.Methods Rattus Mesenchymal Stem Cells were cultured in vitro and 3rd generation cells were induced in 10 μmol/L 5-azacytidine,troponin Ⅰ was checked with immunohistochemistry and Western blot,GATA-4 and Nkx2.5 gene were analyzed by RT-PCR in 1,2,3 and 4 weeks respectively.Results Growth of MSCs stopped after being induced,trending to form colonies,troponin Ⅰ positive expressed since 2nd week,GATA-4 and Nkx2.5 began to express in 1st week,increasing gradually,peak in 3rd week.Conclusion GATA-4 and Nkx2.5 gene expression increased gradually in the course of stem cells differentiation,which may control the rule of cardiomyocyte transcription.更多还原