【摘要】 目的研究淫羊藿苷(icariin,ICA)体外诱导小鼠胚胎干细胞(embryonic stem cells,ESCs)ES-E14细胞分化为心肌细胞的作用。方法复苏的ESCs经直接悬浮法形成拟胚体(EBs),应用ICA定向诱导,相差显微镜下观察分化细胞的形态学变化,免疫细胞荧光技术及Western blot(WB)分别检测心肌细胞特异性肌钙蛋白I(TnI)、心室肌球蛋白轻链(Mlc-1v)的蛋白表达及表达量的变化;透射电镜观察分化细胞的超微结构。结果经ICA诱导后第5天的EBs出现了细胞跳动点,ICA诱导后第3天的细胞心肌细胞特异性肌钙蛋白I(TnI)、心室肌球蛋白轻链(Mlc-1v)的蛋白表达均阳性,于诱导后的第3天(早期)、第12天(中期)及第20天(晚期)心肌细胞特异性肌钙蛋白I(TnI)、心室肌球蛋白轻链(Mlc-1v)的表达逐渐增多,晚期明显高于早期及中期;透射电镜下可见大量平行排列的肌丝。结论用直接悬浮法,ICA体外能诱导鼠ESCs分化为心肌细胞,分化的心肌细胞两种心肌特异性结构蛋白量的表达随分化进程的进展逐渐增多。更多还原

【Abstract】 Objective To study the effect of icarrin on inducing murine embryonic stem(mES) cells to differentiate into cardiomyocyte in vitro. Methods mES cells were cultured as embryoid bodies(EBs) by direct suspension method with icariin. The cardiomyocytes derived from mES cells were characterized by immunofluorescence staining and Western blot for cardiac specific proteins. The ultrastructure of the derived cardiomyocytes was further characterized by transmission electron microscopy. Results Large amount of cardiomyocytes were induced from mES cells by direct suspension culture with icariin treatment. The beating cardiomyocytes derived from mES cells expressed cardiac specific proteins myosin light chain(MLC-1v) and cardiac troponin I(cTnI) . Western blot showed that protein expression levels of MLC-1v and cTnI were all higher in late differentiation stage than in early differentiation stage. Transmission electron microscopy showed obvious structure of cardiomyocytes characterized by the paralleled myofilament. Conclusion Icariin can induce the differentiation of mES cells into beating cardiomyocytes with normal structure.更多还原