【作者】 朱敏洁；

【导师】 唐明；

【作者基本信息】 华中科技大学 ， 生理学， 2008， 博士

【摘要】 目的:研究人参皂甙单体Rg1(Ginsenoside Rg1)在定向诱导分化小鼠D3系胚胎干细胞(D3 embryonic stem cells,D3-ESCs)为心肌样细胞中的作用,以及人参皂甙单体Rg1对D3-ESCs迁移所起的作用。方法:D3-ESCs在基本的细胞培养基中培养,在人参皂甙单体Rg1的定向诱导下,分化为心肌样细胞。用倒置显微镜观察细胞的形态;用免疫细胞化学技术检测特异的α-肌节型肌动蛋白(α-sarcomeric actin);用逆转录PCR技术检测α-心肌肌球蛋白重链的表达(αmyosin heavy chain,α-MHC);用流式细胞技术检测胚胎干细胞的分化率;用迁移滤膜检测胚胎干细胞的迁移率。结果:被人参皂甙单体Rg1诱导后,大多数的D3-ESCs能分化为心肌样细胞,这些细胞中的α-肌节型肌动蛋白(α-sarcomeric actin)染色呈阳性,通过逆转录PCR可以检测到α-心肌肌球蛋白重链的表达(α-MHC)。在D3-ESCs分化前给予人参皂甙Rg1,发现其分化率比对照组高(p < 0.05)。人参皂甙Rg1作用下的D3-ESCs的迁移率比对照组高(p < 0.05)。结论:人参皂甙Rg1(10μmol/L)在体外有促进D3-ESCs诱导分化为心肌样细胞的能力,并且有促进其迁移的能力。更多还原

【Abstract】 Objective: To study the effect of Ginsenoside Rg1 extracorporeal oriented inducing the differentiation of mouse embryonic stem cells(ESCs) into myocardium-like cells and to study the effect of Ginsenoside Rg1 on migraton of ESCs. Methods: Mouse embryonic stem cells were cultured in cell cuture medium and differentiated into myocardium-like cells by oriented induction with Ginsenoside Rg1. The morphology of cells was observed under invert microsceope ; specificα-sarcomeric actin was detected with immunocytochemistry ;α-myosin heavy chain(α-MHC) was examined by RT-PCR ; differentiation ratio of ESCs was detected by flow cytometry ; migration ratio of ESCs was detected by using fluorescence blocking PET track-etched membrane. Results: After being induced by Ginsenoside Rg1, most of ESCs would be differentiated into myocardium-like cells, the positive staining for specificα-sarcomeric actin, the expression ofα-MHC could be detected by RT-PCR. When given Rg1 before ESCs differentiated, ratio of differentiation was higher then control group(p < 0.05). Ratio of migration with Rg1(10μmol/L) was higher then control group(p < 0.05). Conclusion: Ginsenoside Rg1(10μmol/L) have the capacity to promot mouse embryonic stem cells differentiating into myocardium-like cells and migration in vitro.更多还原