心脏营养素-1对骨髓间充质干细胞分化为心肌样细胞的作用  

Effects of Cardiotrophin-1 on Differentiation of Cardiomyocyte-like Cells Induced from Rat Bone Marrow Mesenchymal Stem Cells

作　　者：陈新云[1,2] 曾智[1] 饶莉[1] 李胜富[3] 周斌[4] 陈玉成[1] 

机构地区：[1]四川大学华西医院心内科,成都610041 [2]成都市第一人民医院心内科 [3]四川大学华西医院卫生部移植工程与移植免疫重点实验室 [4]四川大学华西第二医院分子与转化医学实验室

出　　处：《四川大学学报（医学版）》2009年第1期85-88,共4页Journal of Sichuan University(Medical Sciences)

摘　　要：目的观察心脏营养素-1(CT-1)是否能促进经诱导分化剂5-氮杂胞苷(5-aza)诱导的骨髓间充质干细胞(BMMSCs)分化为心肌样细胞,并研究其相关机制。方法自成年大鼠骨髓中分离BMMSCs,分别以普通培养基(A组)、加入含CT-1的培养基(B组)培养、经5-aza诱导后加入普通培养基(C组)及5-aza加入含CT-1的培养基(D组)培养。观察细胞形态的改变,并通过免疫组化分析分化后细胞表达心脏特异性肌钙蛋白T(cTnT)的情况。电镜观察分化后细胞的超微结构及实时荧光定量检测α-actin、β-myosin heavy chain(β-MHC)、Nkx2.5、GATA4基因表达。结果C、D组的BMMSCs在培养4周后均形成心肌样细胞形态,并且均表达cTnT;D组BMMSCs分化的心肌样细胞形成了肌管样结构;D组α-actin、β-MHC、Nkx2.5、GATA4基因表达明显高于C组。结论CT-1可能通过对GATA4、Nkx2.5基因表达的调控而促进5-aza诱导的BMMSCs分化为心肌样细胞。Objective To investigate the effects of cardiotrophin-1 （CT-1） on differentiation of induced rat bone marrow mesenchymal stem cells （BMMSCs） in vitro with 5-azacytidine （5-aza） for the purpose of elucidation of the cellular biological mechanisms. Methods BMMSCs isolated from femur of rats were divided into four groups： untreated group as control （group A）; 0. 1 nmol/L CT-1 added to medium （Group B）; induced with 10 μmol/L 5-aza only （Group C） ; induced with 10 μmol/L 5-aza combined with 0. 1 nmol/L CT-1 added to medium （Group D）. After 4 weeks of induced culturing, the differentiation of induced myocyte like cells were estimated, levels of cardiac troponin-T （cTnT） by immunohistochemical staining and ultrastructure of induce-cultured BMMSCs were determined and mRNA expression of α-actin, β-myosin heavy chain （β-MHC）, Nkx2.5, GATA4 were analyzed by real time polymerase chain reaction （RT-PCR）. Results After 4 weeks of induced culturing, morphological characteristics of myocyte like cells with the expression of cTnT were observed in group C and D cells. Higher level expression of GATA4, Nkx2.5, α- actin and β-MHC mRNA in group D was observed by comparing with those of group C and the differentiated BMMSCs with formations of myofilaments distinctly were also existed in 5-aza combined with CT-1 treatment group. Conclusion This study suggests that induced culturing of BMMSCs in the presence of 5-aza combined with CT-1 can enhance cardiomyocytic characteristics. CT-1 upregulates the expression of GATA4, Nkx2.5, α-actin and β-MHC mRNA, and rapidly promotes the differentiation and maturation of cardiomyocyte-like cells differentiated from BMMSCs induced with 5-aza.

关 键 词：骨髓间充质干细胞 细胞分化 心肌样细胞 心脏营养素-1 

分 类 号：R329[医药卫生—人体解剖和组织胚胎学]