心肌梗死大鼠血清促进大鼠骨髓间质干细胞分化为心肌细胞的作用

Effect of serum from acute myocardial infarction rat on inducing rat bone marrow mesenchymal stem cells into cardiomyocytes

目的:探讨急性心肌梗死(AMI)大鼠血清在体外对大鼠骨髓间质干细胞(BMSC)分化为心肌细胞的作用。方法:将大鼠BMSC分为6组进行细胞培养:Ⅰ组为未诱导组(对照组),Ⅱ组为5-aza诱导组,Ⅲ组为5-aza+AMI血清诱导组,Ⅳ组为5-aza+正常大鼠血清诱导组,Ⅴ组为AMI血清诱导组,Ⅵ组为正常大鼠血清诱导组。培养30d后进行分化细胞鉴定,检测细胞的形态变化和心肌肌钙蛋白(cTnT)、心肌细胞转录因子GATA-4和结蛋白(desmin)的表达。结果:大鼠BMSC经5-aza或5-aza联合血清诱导后,向心肌样细胞分化,其cTnT、GATA-4、desmin表达阳性。对照组和单纯血清诱导组的BMSC未见形态改变,其cTnT亦为阴性,但GATA-4、desmin却呈弱阳性。另外,5-aza联合血清诱导组在诱导2周后可观察到部分细胞呈缓慢而有节律的跳动,而单纯5-aza诱导组未见细胞跳动。结论:单纯用AMI大鼠血清不能诱导BMSC向心肌细胞分化,但AMI血清能促进5-aza诱导的BMSC向心肌细胞分化,并促进分化的心肌细胞成熟。

AIM： To induce rat bone marrow mesenchymal stem cells （BMSC） into cardiomyocytes and investigate the influence of serum coming from acute myocardial infarction （AMI） rat on the procedure. METHODS ： The passage 3 BMSC were divided into six groups： group Ⅰwas control group; group Ⅱ was induced with 5 -azacytidine; group Ⅲwas induced with 5 - azacytidine and serumfrom AMI rat; group Ⅳ was induced with 5 - azacytidine and serum from normal rat; groupⅤ and group Ⅵwere induced with serum from AMI rat or normal rat. The cardiac troponin T, GATA -4 and desmin were detected 30 days after induction. RESULTS ： After inducing by 5 - azacytidine, 5 - azacytidine and two kinds of serum, some cells in the three groups differentiated into cardiac like cells. The expressions of cardiac troponin T, GATA-4 and desmin were positive in cells differentiated from BMSC. The troponin T expression in control group and group inducing by AMI serum alone were negative but GATA -4 and desmin expressed weakly. Some cells induced with 5 - azacytidine and serum were slowly beating 2 weeks after induction, but the cells induced with 5 - azacytidine alone was not beating. CONCLUSION： Serum from AMI can not induce BMSC to differentiate into cardiomyocytes, but it promotes BMSC differentiate into cardiomyocytes induced by 5 - azacytidine and facilitate the differentiated cells to mature.