多柔比星、二甲双胍对干细胞心肌样分化的代谢组分研究  

Effects of doxorubicin and metformin on metabolic componentsof differentiation of stem cells into cardiomyocyte-like cells

作　　者：邸志权[1] 胡金芳[1] 姜一朴 安梦培 付合明 冯玥[1] 申秀萍[1] DI Zhi-quan;HU Jin-fang;JIANG Yi-pu;AN Meng-pei;FU He-ming;FENG Yue;SHEN Xiu-ping(Tianjin Institute of Pharmaceutical Research New Drug Assessment Co.Ltd,Tianjin 300301,China;College of Pharmacy,Anhui Medical University,Hefei 230032,China)

机构地区：[1]天津药物研究院新药评价有限公司,天津300301 [2]安徽医科大学药学院,安徽合肥230032

出　　处：《中国药理学通报》2018年第11期1560-1566,共7页Chinese Pharmacological Bulletin

基　　金：国家科技重大专项(No 2015ZX09501004-002-003);天津市科技计划项目(No 16PTGCCX00090;17ZXXYSY00020)

摘　　要：目的结合代谢组学技术和胚胎干细胞心肌定向诱导分化方法,对多柔比星和二甲双胍造成代谢组分的变化进行研究。方法以小鼠胚胎干细胞体外心肌定向诱导分化为模型,分化培养液中添加不同剂量的多柔比星(3.40、0.17μmol·L^(-1))以及二甲双胍(905.7μmol·L^(-1))。通过对诱导形成拟胚体的一般观察,统计自主节律跳动、拟胚体的数量、频率等;免疫细胞化学法检测心肌特异性蛋白—心肌肌钙蛋白T(cTnT)的表达;应用代谢组学技术初步筛选差异代谢图谱,以多柔比星和二甲双胍共同影响的代谢组分为目标,尝试寻找心脏毒性敏感标志物。结果一般观察和免疫细胞化学结果显示,多柔比星高浓度明显抑制向心肌细胞分化,cTnT表达明显增加,低浓度组拟胚体自主节律明显增快,cTnT表达变化不明显;二甲双胍未见明显变化。代谢组学结果显示,多柔比星和二甲双胍可致细胞内同型半胱氨酸(Hcy)、铁卟啉(FeTMPyP)、叶酸、半胱氨酸(Cys)、cAMP等组分明显降低。结论氧化应激在心脏分化发育过程中起重要作用,Hcy、FeTMPyP、Cys等具有成为心脏毒性敏感标志物的潜力。Aim To analyze the changes of the metabolic profiling of cardiomyocyte-like cells via introducing metabonomics and stem cell differentiation tech-nology.Methods The mouse embryonic stem cells(mESCs)were induced to differentiate into cardiomyocytes in vitro,then different doses of doxorubicin(3.40,0.17μmol·L^-1)and metformin(905.7μmol·L^-1)were added into the differentiation mediums.Through general observation of the development of the embryoid bodies,the numbers and frequencies of the autonomic rhythmic embryoid bodies were counted.The cardiac specific troponin protein T(cTnT)expression was detected by immunocytochemistry.Finally,some sensitive markers for cardiac toxicity were tried to find in the metabolic components affected by both doxorubicin and metformin together.Results The results of general observation and immunocytochemistry showed that doxorubicin 3.40μmol·L^-1 significantly inhibited the differentiation of mESCs,and the expression of cTnT increased significantly,but the spontaneous rhythms of the embryos increased significantly in DOX 0.17μmol·L^-1,and the expression of cTnT remained unchanged.Besides,metformin did not change significantly.Metabonomics results showed that homocysteine(Hcy),iron porphyrin(FeTMPyP),folic acid,cysteine(Cys),cAMP and other components were down-regulated by doxorubicin and metformin even at low concentration.Conclusions The oxidative stress plays a vital role in the process of cardiac differentiation and development.Hcy,FeTMPyP,Cys and so on have the potential as sensitive markers of cardiac toxicity.

关 键 词：胚胎干细胞 心肌细胞 代谢组学 多柔比星 二甲双胍 诱导分化 

分 类 号：R322.11[医药卫生—人体解剖和组织胚胎学] R329.24[医药卫生—基础医学]