5-氮杂胞嘧啶诱导人骨髓基质干细胞心肌分化  

Myocardial Differentiation of Human Bone Marrow Stromal Cells Induced by 5-Azacytidine

作　　者：胡昕婴[1] 史剑慧[1] 牛玉宏[1] 葛均波[1] 

机构地区：[1]复旦大学干细胞研究中心,上海市心血管病研究所,复旦大学附属中山医院心内科,上海200032

出　　处：《中国临床医学》2004年第3期298-299,共2页Chinese Journal of Clinical Medicine

基　　金：上海市医学发展基金 -重点研究课题基金 ( 2 0 0 0I -2D0 0 2 );上海市科委优秀学科带头人计划资助项目 ( 0 3XD 14 0 10 )

摘　　要：目的 :研究 5 -氮杂胞嘧啶 (5 -aza)诱导人骨髓基质干细胞 (hMSC)心肌分化过程中心肌特异转录因子和心肌特异基因的表达变化。方法 :取传至第 8代hMSC ,以 3μmol/L的 5 -aza孵育细胞 2 4h ,分别在诱导后 3d、1周、2周和 3周 4个时间点提取细胞RNA ,以RT -PCR方法检测细胞心肌特异转录因子Nkx2 .5和心肌特异基因 (connexin4 3、ANP等 )的表达。结果 :hMSC经 5 -aza诱导后 ,形成肌节样结构。RT -PCR的结果显示 ,5 -aza可诱导心肌特异基因connexin4 3和ANP的表达 ,随诱导时间延长 ,connexin4 3和ANP的表达量逐渐增加。 5 -aza还能诱导hMSC表达心肌特异转录因子Nkx2 .5 ,并持续表达至 3周。结论 :5 -aza在体外可诱导hMSC向心肌细胞分化 ,5 -aza可能通过诱导心肌特异转录因子的表达而启动hMSC心肌化的过程。Objective: We investigated the potential and temporal transcription regulation of human bone marrow stromal cells(hMSC) being induced to cardiomyocytes in vitro with the treatment of 5-azacytidine(5-aza). Methods: Cardiomyocyte differentiation was induced by the treatment of 3μmol/L of 5-aza. We investigated the morphological change after the induction. 3 days, 1 week, 2 weeks, 3 weeks after being induced by 5-aza, the expression of cardiomyocyte-specific genes connexin43, atrial natriuretic protein (ANP) and cardiomyocyte-specific transcription factors Nkx2.5 in hMSC were assessed using RT-PCR assay to investigate the temporal regulation of cardiomyogenesis at a transcriptional level. Results: After the treatment of 5-aza, the former fibrobalst-like cells inclined to be unidirectioned and formed a sarcomere-like structure. RT-PCR assessment showed that the differentiated cells began to express Nkx2.5, one of important cardiac transcript factors, connexin43 and ANP, the specific cardiac genes, which could last at least 3 weeks. Conclusion:These findings show that hMSC possesses the differentiation potential of cardiomyocyte. We demonstrate that cardiomyocyte differentiation of hMSC could be induced by 5-aza in vitro. The reagent functioned at the transcriptional level to promote cardiomyogenesis of hMSC.

关 键 词：5-氮杂胞嘧啶 人骨髓基质干细胞 细胞分化 心肌细胞 CONNEXIN43 ANP 

分 类 号：R329.2[医药卫生—人体解剖和组织胚胎学]