体外诱导骨髓间充质干细胞向心肌细胞分化前后离子通道基因表达的变化  

Changes of ion channel mRNA expression in bone marrow mesenchymal stem cells before and after differentiation into cardiomyocytes in vitro

作　　者：时飒[1] 金连弘[1] 刘慧文[1] 申景岭[1] 李弘[2] 徐长庆[2] 

机构地区：[1]哈尔滨医科大学组织与胚胎学教研室,黑龙江省哈尔滨市150086 [2]哈尔滨医科大学病理生理教研室,黑龙江省哈尔滨市150086

出　　处：《中国临床康复》2006年第17期42-44,i0002,共4页Chinese Journal of Clinical Rehabilitation

摘　　要：目的:观察体外诱导骨髓间充质干细胞向心肌细胞分化前后某些离子通道mRNA表达的变化,以进一步探讨离子通道在骨髓间充质干细胞增殖和向心肌细胞分化中的作用。方法:实验于2004-12/2005-07在哈尔滨医科大学病理生理学实验室和组织与胚胎学实验室完成。实验选用Wistar大鼠,雌雄不限,体质量100～120g。分离、培养和纯化大鼠骨髓间充质干细胞,以DAPI(5mg/L)标记第三代的骨髓间充质干细胞后,采用10μmol/L5-氮胞苷诱导,于诱导后第4周做细胞免疫荧光化学检测细胞肌钙蛋白的表达。采用反转录聚合酶链反应技术检测诱导分化前后钙通道的α1c,α1D,α1G,α1H,α1S亚基;钾通道:ERG,IK1,KvLQT1和钠通道SCN5AmRNA的表达结果:①5-氮胞苷诱导后第4周细胞可见心肌细胞特异性肌钙蛋白的表达。②反转录聚合酶链反应结果显示分化后的细胞表达L-型钙通道的α1c,α1D,T-型钙通道的1G亚基和钾通道:ERG-1,Kir2.1,KvLQT-1和钠通道SCN5AmRNA的表达,与成熟心肌细胞相似。③在诱导分化前的骨髓间充质干细胞也发现了L-型钙通道的α1c亚基,ERG-1和KvLQT-1钾通道的表达。结论:骨髓间充质干细胞可被诱导分化为心肌样细胞,分化后的细胞表达形成动作电位的相关离子通道基因,进一步表明骨髓间充质干细胞可作为心肌细胞移植的种子细胞。实验观察到未分化的骨髓间充质干细胞也有L-型钙通道,ERG-1和KvLQT1钾通道的表达,推测这些离子通道可能在骨髓间充质干细胞的增殖和向心肌细胞分化中具有重要的作用。ATM： To investigate the mRNA expression of ion channel in bone marrow mesenchymal stem cells （MSCs） cultured in vitro before and after differentiation into cardiomyocytes, so that we can further probe into the effect of ion channel in the proliferation of MSCs and differentiation into cardiomyocytes. METHODS： This experiment was conducted at the Laboratory of Pathopsychology and Laboratory of Histology and Embryology, Harbin Medical University from December 2004 to July 2005. Wistar rats of either gender, with body mass of 100 to 120 g, were used in this experiment. MSCs were isolated, cultured and purified, and MSCs of the third generation were labeled with DAPI （5 mg/L）. MSCs were induced with 10 μmol/L 5-azaeytidine to differentiate into eardiomyocytes. The expression of cellular troponin was detected by immunofluorescent staining 4 weeks after induction. Reverse transcription-polymerase chain reaction （RT-PCR） was used to detect the mRNA expression of α1c,α1D,α1G, α1H,α1S subunit of calcium channel and ERG,IKI,KVLQTI of potassium channels and SCNSA of sodium channel. RESULTS： ①The expression of specific treponin of cardiomyocytes could be seen 4 weeks after the induction of 5- azacytidine. ②The results of RT-PCR showed that the mRNA of α1c,α1D expressed by differentiated cells for L-type calcium channel , IG subunit for T-type calcium channel, ERG-1 ,Kir2.1 ,KVLQT-1 for potassium channels and SCNSA for sodium channel were similar to the mature cytomyocytes. ③The expression of α1c subunit , and ERG-1 and KVLQT-1 of potassium channels could also be found in the MSCs before induction and differentiation. CONCLUSION： MSCs could be induced and differentiated into eardiomyocytes . The mRNA of some ion channels, which produced action potential, was expressed in the differentiated cells. It is further demonstrated that BMCs could be used as seed cells of eardiomyocyte transplantation. The expression of L-type calcium channel, ERG-1 and KVLQTI of potassium channels could also b

关 键 词：干细胞 心肌 细胞分化 离子通道 基因表达 

分 类 号：R394.2[医药卫生—医学遗传学]