Effect of endothelin-1 on differentiation of cardiomyogenic cells induced from rabbit bone mesenchymal stem cells / 中国病理生理杂志

ABSTRACT

AIM: To investigate biological effect of endothelin-1 (ET-1) on differentiation of cardiomyogenic cells (CGCs) induced from rabbit bone mesenchymal stem cell (BMSCs) in vitro. METHODS: The BMSCs from shaft of femur of Japan rabbits were isolated and propagated for cell culture. Cultured BMSCs were randomly divided into six groups Group Ⅰ the control group (uninduced group); Group Ⅱ the ET-1 group, added ET-1 (30 nmol/L) into culture medium; Group Ⅲ 5-aza group, added 5-aza (10 ?mol/L) into culture medium; Group Ⅳ 5-aza+ET-1 group, after inducing with 5-aza for 3 weeks, ET-1 was added into culture medium. This group was divided into three parts-Ⅳ 1、Ⅳ 2、Ⅳ 3, and separated to add 10、 30、 50 nmol/L ET-1. During the cell culture, growing and differentiation of BMSCs were observed. After inducing for 4 weeks, the differentiation rate and the diameter of cardiomyogenic cells were calculated; Western-blot was employed to analyze the expressions of GATA-4 protein and phosphorylation level. The expression of ?-MHC mRNA was assessed by RT-PCR. Immunohistochemistry staining of Troponin-I and ultrastructure observation of induced cardiomyogenic cells were also completed simultaneously. RESULTS: The cell diameters of CGCs in group Ⅳ 2 were enlarged significantly (P0.05). In group Ⅲ and Ⅳ 2 , the positive cells of cTroponin I staining were more, the expression of ?-MHC mRNA was significantly increased (P