文献类型：期刊文献

中文题名：黄芪甲苷诱导大鼠骨髓间充质干细胞向心肌样细胞分化的研究

英文题名：Astragaloside Effect on the Differentiation of Rat Bone Marrow Mesenchymal Stem Cells into Cardiomyogenic Cells in Vitro

作者：赵静苗[1];胡继宏[1];王秋萍[1]

第一作者：赵静苗

机构：[1]甘肃中医药大学甘肃重大疾病分子医学与中医药防治研究省级重点实验室,甘肃省兰州市730000

第一机构：甘肃中医药大学科研实验中心（甘肃省中医药标准化技术委员会秘书处）

年份：2017

卷号：20

期号：21

起止页码：2635

中文期刊名：中国全科医学

外文期刊名：Chinese General Practice

收录：CSTPCD;;Scopus;北大核心:【北大核心2014】；

基金：国家自然科学基金资助项目(81260597)

语种：中文

中文关键词：肌细胞,心脏;细胞分化;骨髓间充质干细胞;黄芪甲苷

外文关键词：Myocytes; cardiac; Cell differentiation; Bone marrow mesenchymal stem cells; Astragaloside

摘要：目的探讨黄芪甲苷(AST)诱导SD大鼠骨髓间充质干细胞(BMSCs)向心肌样细胞分化的作用。方法对第2代BMSCs进行传代培养,采用四甲基偶氮唑蓝(MTT)法检测不同浓度AST在24、48、72 h时对BMSCs的增殖作用。取第4代BMSCs分为BMSCs组、5-氮杂胞苷组(5-AZA组)和AST组,5-AZA组和AST组分别加入浓度为10μmol/L的5-AZA和浓度为100μmol/L的AST诱导液进行诱导,采用荧光倒置相差显微镜观察诱导后各组细胞的形态变化。通过Real-time q PCR法检测诱导后各组结蛋白(Desmin)、α横纹肌肌动蛋白(α-SCA)、肌钙蛋白I(cTnI)mRNA表达水平;采用Western blotting法检测各组肌钙蛋白T(cTnT)表达水平。结果 100μmol/L AST亚组孔72 h时BMSCs增殖情况好于24、48 h时及其他浓度AST亚组孔各时间点(P<0.05)。荧光倒置相差显微镜观察显示AST体外诱导SD大鼠BMSCs形态分化类似心肌样细胞。5-AZA组、AST组Desmin、α-SCA、cTnI mRNA表达水平表达高于BMSCs组(P<0.05),且AST组以上蛋白mRNA表达水平高于5-AZA组(P<0.05)。与BMSCs组组比较,5-AZA组和AST组cTnT表达水平升高(P<0.05);与5-AZA组比较,AST组cTnT表达水平下降(P<0.05)。结论浓度为100μmol/L的AST作用72 h时能够在体外诱导BMSCs向心肌样细胞分化,但分化效果弱于5-AZA。
Objective To investigate the astragaloside（ AST） effect on the differentiation of bone marrow mesenchymal stem cells（ BMSCs） into cardiomyogenic cells in SD rats. Methods BMSCs were subcultured on behalf of BMSCs. The proliferation of BMSCs was detected by MTT assay at 24,48,72 h after the inducing with different concentrations of AST. The 4th generation of BMSCs were divided into BMSCs group,5-azacytidine group（ 5-AZA group） and AST group,cultured with the culture medium with 10% fetal bovine serum,the culture medium with 10 μmol/L 5-AZA,and the culture medium with 100 μmol/L AST, respectively. Morphological changes of BMSCs were observed by phase contrast fluorescence inverted microscopy. The expression levels of Desmin mRNA,α-Sarcomeric actin（ α-SCA） mRNA and troponin（ c Tn I）mRNA in the BMSCs group,5-AZA group and AST group were measured by Real-time q PCR,and the expression levels of c Tn T in these three groups were detected by western blotting. Results The proliferation of BMSCs was the best when they were induced for 72 h by AST at the concentration of 100 μmol/L（ P〈0. 05）. Morphological differentiation of BMSCs induced by AST in vitro was similar to that of cardiomyogenic cells. The BMSCs group had lower expression levels of Desmin mRNA,α-SCA mRNA and c Tn I mRNA than 5-AZA group and AST group（ P〈0. 05）. Compared with 5-AZA group,the expression levels of Desmin mRNA,α-SCA mRNA and c Tn I mRNA in AST group were higher（ P〈0. 05）. 5-AZA group and AST group had higher c Tn T expression levels than the BMSCs group（ P〈0. 05）. Compared with 5-AZA group, AST group had lower expression levels of c Tn T（ P〈0. 05）. Conclusion The differentiation of BMSCs into cardiomyogenic cells in vitro can be promoted after being induced by AST with a concentration of 100 μmol/L for 72 hours,but the induction effect of AST is weaker than 5-AZA.