来自:中国糖尿病杂志 编辑: 魏蕊 杨进 洪天配 高美娟 侯文芳 刘|点击数:|2014-08-28
【关键词】Ghrelin;人胚胎干细胞;心肌细胞;分化
【Abstract】Objective To investigate the effects of ghrelin on cardiomyocyte differentiation of human embryonic stem (hES) cells. Methods The hES cells were differentiated into cardiomyocytes with or without different concentrations of ghrelin. The appearance of spontaneously contracting clusters was counted microscopically and the expression of cardiomyocyte-specific markers was determined through real-time PCR. The expression of growth hormone secretagogue receptor (GHS-R) 1α was detected by RT-PCR, Western blot and Immunostaining. Furthermore, GHS-R1α antagonist [D-lys3]-GHRP-6 was used to determine whether GHS-R1α was involved in the ghrelin-induced cardiomyocyte differentiation. Results Compared with control group, the proportion of spontaneously contracting clusters was increased by 10-10, 10-9 and 10-8 mol/L of ghrelin [percentages of beating EBs were (12.9±1.4)%, (19.5±2.2)% and (13.5±3.1)% vs (10.3±2.2)% respectively, P<0.05]. RT-PCR analysis showed that the mRNA levels of α-myosin heavy chain (α-MHC) and cardiac troponin I (cTnI) were upregulated in ghrelin-treated group [α-MHC: 100% vs (40.1±13.2)%, P<0.001; cTnI: 100% vs (52.6±9.8)%, P<0.001]. RT-PCR, Western blot and immunostaining analyses revealed that GHS-R1α was detected in the differentiated cells at different stages
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