中药麦冬对骨髓间充质干细胞分化心肌样细胞的诱导作用研究

目的研究为证明浙麦冬可能具有MSCs诱导作用,且其补心阴作用与骨髓间充质干细胞(bone marrow mesenchymalstem cells,MSCs)转化率成正比。且阐明浙麦冬对MSCs的诱导作用和机制。方法(1)流式细胞仪鉴定大鼠骨髓MSCs的表面标记分子CD73,CD90,CD45,CD34,观察药物对SD大鼠MSCs诱导分化心肌样细胞形态结构的影响。(2)免疫荧光法检测浙麦冬对SD大鼠MSCs诱导分化心肌样细胞中CTnI和CX43蛋白表达情况。(3)qPCR法检测MSCs诱导分化心肌样细胞中NKx 2.5,GATA4和β-MHC mRNA表达情况。结果(1)与空白对照组比较,给药组的心肌样细胞中肌钙蛋白I(CTnI)和连接蛋白43(CX43)的表达量明显增加,而且呈现剂量依赖性和培养时间依赖性。(2)与空白对照组比较,给药组的心肌样细胞中NKx 2.5,GATA4和β-MHC mRNA相对表达量明显增加,而且呈现剂量依赖性和培养时间依赖性。(3)与空白对照组比较,给药组的MSCs中在1周左右存活的部分细胞体积逐渐增大,呈球状、棒状或长梭形;2周以后细胞间出现连接,排列方向渐趋一致。结论补心阴中药浙麦冬能诱导MSCs分化形成心肌样细胞,而且呈现剂量依赖性。

Effect of Ophiopogon Japonicus Inducing Differentiation of Bone Marrow Mesenchymal Stem Cells in Myocardial Cells

Objective This study is to prove that Ophiopogon japonicus may have MSCs-inducing effect,and the nourishing heart Yin effect is directly proportional to the conversion rate of MSCs.And it is to elucidate the induction and mechanisms of MSCs in Ophiopogon japonicus.Methods(1)Flow cytometry identified the surface marker including CD73,CD90,CD45 and CD34 of rat bone marrow MSCs and we observed the effects of drugs on the morphology and structure of myocardial cells induced by MSCs in SD rats.(2)Immunofluorescence assayed the expressions of CTnI and CX43 protein in myocardial cells induced by MSCs in SD rats.(3)The qPCR method assayed the expressions of NKx 2.5,GATA4 andβ-MHC mRNA in differentiated myocardial cells induced by MSCs.Results(1)Compared with the blank control group,the expression levels of cardiac troponin I(CTnI)and connexin 43(CX43)in the myocardial cells of the medication administration team were significantly increased in a dose-dependent and culture-dependent manner.(2)Compared with the blank control group,the relative expressions of NKx 2.5,GATA4 andβ-MHC mRNA in the myocardial cells of the medication administration team were significantly increased in a dose-dependent and culture-dependent manner.(3)Compared with the blank control group,the volume of some cells that survived in the MSCs of the medication administration team increased gradually in a week,and it was spherical,rod-shaped or long fusiform.After 2 weeks,the cells appeared to be connected,and the arrangement direction became more consistent.Conclusion The heart Yin-nourishing Chinese medicine Ophiopogon japonicas can induce differentiation of bone marrow mesenchymal stem cells in myocardial cells in a dose-dependent manner.