低剂量乌头碱持续给药重塑hiPSCs-CM线粒体能量代谢模式  

Repeated administration of low-dose aconitine remodels mitochondrial energy metabolism pattern of human iPSCs-CM

作　　者：刘泓 岳兰昕 邱丽珍[3] 肖成荣 王淑美 周维 高月 LIU Hong;YUE Lan-xin;QIU Li-zhen;XIAO Cheng-rong;WANG Shu-mei;ZHOU Wei;GAO Yue(School of Traditional Chinese Medicine Guangdong Pharmaceutical University,Guangzhou 510006,China;Institute of Radiation Medicine,Academy of Military Medical Sciences,Beijing 100850,China;Academy of Traditional Chinese Medicine Tianjin University of Traditional Chinese Medicine,Tianjin 301617,China)

机构地区：[1]广东药科大学中药学院,广东广州510006 [2]军事科学院军事医学研究院辐射医学研究所,北京100850 [3]天津中医药大学中医药研究院,天津301617

出　　处：《中国药理学通报》2021年第5期617-623,共7页Chinese Pharmacological Bulletin

基　　金：国家自然科学基金资助项目(No 8180140316,8163000131)。

摘　　要：目的探究低剂量乌头碱对人诱导多能干细胞分化心肌细胞(human induced pluripotent stem cell-derived cardiomyocytes,hiPSCs-CM)代谢的影响。方法100 nmol·L^(-1)乌头碱对hiPSCs诱导分化的hiPSCs-CM细胞持续给药后,能量代谢分析仪(Seahorse)检测代谢表型变化;微电极阵列(Microelectrode array,MEA)检测电生理参数;qRT-PCR检测脂糖代谢关键基因的表达;Western blot检测hiPSCs-CM中多能性转录因子的表达。结果100 nmol·L^(-1)乌头碱持续给药后,hiPSCs-CM代谢表型从氧化磷酸化向糖酵解方向偏移;脂代谢调控关键基因表达降低,而葡萄糖转运蛋白1(facilitative glucose transporters 1,Glut1)与转运蛋白4(facilitative glucose transporters 4,Glut4)比值明显升高;hiPSCs-CM收缩力降低(收缩频率增加且振幅减弱)、兴奋传导延迟,但未见明显心律失常指征。结论低剂量乌头碱持续给药会诱导心肌功能出现明显变化、能量代谢模式发生明显重构,这可能与其导致心肌细胞多能性增强和/或能量代谢底物选择改变密切相关。Aim To study the effect of low dose aconitine on the metabolism of hiPSCs-CM.Methods After 100 nmol·L^(-1) aconitine continuously administered in hiPSCs-CM differentiated from hiPSCs,seahorse was used to detect the changes in the metabolic phenotype of hiPSCs-CM;microelectrode array(MEA)was used to detect electrophysiological parameters of hiPSCs-CM;qRT-PCR was used to detect the expression of key genes regulating in lipid metabolism and glucose metabolism;Western blot was used to detect the expression of stem cell pluripotency regulator factors.Results After continuously treated with low dose aconitine,the metabolism of hiPSCs-CM shifted from oxidative phosphorylation to glycolysis.The expression of key genes involved in lipid metabolism decreased,and the facilitative glucose transporters 1(Glut1)/facilitative glucose transporters 4(Glut4)ratio also significantly decreased.The contractility of hiPSCs-CM was reduced(increased contractile frequency and decreased beat amplitude)and excitatory conduction delayed,but no obvious indications of arrhythmia were observed.Conclusions Long-term administration of low dose aconitine induces significant changes in myocardial function,and the energy metabolism patterns are also significantly remodeled,which may be related to the enhancement of pluripotency of cardiomyocytes and/or the change in the selection of substrates for energy metabolism caused by long-term administration of low dose aconitine.

关 键 词：乌头碱 低剂量 hiPSCs-CM 能量代谢 氧化磷酸化 糖酵解 

分 类 号：R-332[医药卫生] R284.1