bFGF和IGF-1对骨髓间充质干细胞向心肌样细胞分化促进作用及相关机制探讨  

bFGF and IGF-1 promotes differentiation of bone mesenchymal stem cells into cardiomyocyte-like cells and its related mechanisms

作　　者：王巧敏 刘洋[1] 吕洋[1] 郭蕊 高京 马聪聪 王海萍[1,2] WANG Qiao-min;LIU Yang;LYU Yang;GUO Rui;GAO Jing;MA Cong-cong;WANG Hai-ping(Dept of Histology and Embryology,Basic Medical College,Hebei North University,Zhangjiakou,Hebei 075000,China;Laboratory of Stem Cell and Reproductive Biology,Hebei North University,Zhangjiakou,Hebei 075000,China)

机构地区：[1]河北北方学院基础医学院组织学与胚胎学教研室,河北张家口075000 [2]河北北方学院干细胞与生殖生物学实验室,河北张家口075000

出　　处：《中国药理学通报》2023年第4期715-722,共8页Chinese Pharmacological Bulletin

基　　金：河北省自然科学基金资助项目(No C2019405091);河北省高等学校科学技术研究项目(No ZD2019066)。

摘　　要：目的 探讨成纤维细胞生长因子(basic fibroblast growth factor, bFGF)和胰岛素样生长因子-1(insulin-like growth factor-1,IGF-1)单独或联合使用对骨髓间充质干细胞(bone mesenchymal stem cells, BMSCs)向心肌细胞(cardiomyocytes, CMs)分化的影响,以及IGF-1和bFGF促分化的作用机制。方法 IGF-1和bFGF单独或联合使用,以及IGF-1和bFGF分别与LY294002联用,诱导4周后,采用免疫细胞化学、免疫荧光检测心肌特异性蛋白表达水平;Western blot检测心肌特异性蛋白和通路相关蛋白表达水平;透射电镜观察细胞的超微结构。诱导1、2和4周后,采用RT-qPCR检测心肌转录因子表达水平。结果 IGF-1和bFGF单独或联合组,均提高心肌特异性蛋白和基因表达水平,且最高表达水平出现在IGF-1和bFGF联合组。IGF-1和bFGF联用组,透射电镜可观察到平行排列的肌丝,以及线粒体和内质网等丰富细胞器。IGF-1和bFGF分别与LY294002联用组,相较于IGF-1和bFGF单独诱导组降低心肌特异性蛋白和基因的表达水平,降低通路相关蛋白Akt磷酸化水平。结论 IGF-1和bFGF均可诱导BMSCs向CMs的定向分化,且两种生长因子的联合是一种更加高效的诱导方法。PI3K/Akt信号通路参与了IGF-1和b FGF促进BMSCs向CMs分化的过程。Aim To elucidate the biological effects of insulin-like growth factor-1(IGF-1)and basic fibroblast growth factor(bFGF)alone or in combination on the differentiation of bone mesenchymal stem cells(BMSCs)into cardiomyocytes(CMs),and to explore the mechanism in the differentiation process of BMSCs into CMs induced by IGF-1 or bFGF.Methods After four weeks of BMSCs induced by induction differentiation medium(with or without LY294002)containing IGF-1 and/or bFGF,the expression levels of proteins associated with the cardiomyogenic phenotype in BMSCs were detected via immunocytochemistry,immunofluorescence staining,and Western blot.Meanwhile,the expression levels of pathway related proteins were detected by Western blot.The cell ultrastructure was observed by transmission electron microscopy(TEM).The expression levels of myocardial specific genes were measured via RT-qPCR.Results Compared with the control group,the expression levels of myocardial specific proteins and genes significantly increased in the IGF-1,bFGF and combination groups and were the highest in the coinduction group.The TEM of the coinduction group showed parallel myofilaments,mitochondria,endoplasmic reticulum and so on.The additon of LY294002 decreased the expression levels of myocardial specific proteins,genes and phosphorylation Akt.Conclusions The effect of IGF-1 combined with bFGF on the differentiation of BMSCs into CMs is markedly better than that induced by IGF-1 or bFGF alone,and the differentiation process may depend on the PI3K/Akt signaling pathway.

关 键 词：成纤维细胞生长因子 骨髓间充质干细胞 心肌细胞 胰岛素样生长因子-1 心肌梗死 PI3K/Akt 

分 类 号：R-332[医药卫生] R322.11R329.24R329.411R542.22R977.6