人胚胎干细胞分化的心肌细胞心脏毒性评价模型的建立及应用

发布时间：2018-07-03 09:13

本文选题：人胚胎干细胞分化的心肌细胞 + 心脏毒性　；参考：《中国新药杂志》2017年07期

【摘要】：目的:建立适合于药物心脏毒性评价的人胚胎干细胞分化的心肌细胞(hESC-CM)模型,并采用此模型评价化合物潜在的心脏毒性。方法:研究选用不同种类的心脏毒性药物:蒽环类抗癌药阿霉素(0.2,1,5,10和20μmol·L~(-1))、酪氨酸激酶抑制剂伊马替尼(0.2,1,5,10和20μmol·L~(-1))、β受体激动剂异丙肾上腺素(0.001,0.01,0.1,1和10μmol·L~(-1))、钙离子通道阻滞剂维拉帕米(0.1,0.3,1,3和10μmol·L~(-1)),分别与hESC-CM孵育,采用实时细胞分析法和高内涵细胞成像技术实时监测心肌细胞搏动的频率、振幅、规则性及线粒体膜电位等指标在给药前后的变化,建立人胚胎干细胞分化的心肌细胞体外心脏毒性评价模型。应用此模型评价化合物A的心脏毒性。结果:阿霉素各浓度与hESC-CM共同孵育时,细胞的搏动振幅、搏动频率、搏动规则指数以及线粒体膜电位均降低;20μmol·L~(-1)的伊马替尼抑制hESC-CM的搏动振幅、搏动频率、搏动规则指数和线粒体膜电位;异丙肾上腺素降低hESC-CM的搏动振幅、升高其搏动频率,但此作用具可逆性,对搏动规则指数、线粒体膜电位无明显影响;维拉帕米迅速降低hESC-CM的搏动振幅、搏动频率,但此作用具可逆性,细胞的搏动规则指数、线粒体膜电位无明显变化。测试化合物A和其阳性对照药托法替尼对hESC-CM的影响类似,对搏动振幅的抑制作用成浓度时间相关性,对搏动频率、搏动频率规则指数也有明显的作用,对线粒体膜电位的影响不显著。结论:利用人胚胎干细胞分化的心肌细胞可以建立一种可行的体外心脏毒性评价模型,结合实时细胞分析法以及高内涵细胞成像技术可快速、灵敏、准确的评价药物的心脏毒性。
[Abstract]:Aim: to establish a cardiomyocyte model (hESC-CM) suitable for evaluating the cardiotoxicity of human embryonic stem cells and to evaluate the potential cardiac toxicity of the compounds. Methods: different kinds of cardiotoxic drugs were selected: adriamycin (0.2 mol L-1) and imatinib (0.2 渭 mol L ~ (-1), tyrosine kinase inhibitor) and isoprenaline (0.0010.010.1kan1 and 10 渭 mol L-1), calcium channel blockers), 20 渭 mol L ~ (-1), 尾 agonist (0.0010.01) and 10 渭 mol L ~ (-1), calcium channel blockers, respectively. Parmi (0.1 ~ 0.3L ~ (-1),) and 10 渭 mol / L ~ (-1), were incubated with hESC-CM, respectively. The changes of cardiomyocyte pulsation frequency, amplitude, regularity and mitochondrial membrane potential before and after administration were monitored by real-time cell analysis and high intension cell imaging. To establish a cardiotoxicity evaluation model of human embryonic stem cell differentiation in vitro. The cardiac toxicity of compound A was evaluated by this model. Results: when incubated with hESC-CM at various concentrations of doxorubicin, the pulsatile amplitude, pulsatile frequency, pulsatile regular index and mitochondrial membrane potential of hESC-CM were decreased by imatinib at 20 渭 mol L ~ (-1). The pulsatile regular index and mitochondrial membrane potential, isoproterenol decreased the pulsatile amplitude of hESC-CM and increased its pulsatile frequency, but the reversibility of this instrument had no significant effect on the pulsatile regular index and mitochondrial membrane potential. Verapamil decreased the pulsatile amplitude and frequency of hESC-CM rapidly, but the reversibility of the apparatus, the regular index of cell pulsation, and the mitochondrial membrane potential did not change significantly. The effect of Tofatinib on hESC-CM was similar to that of the positive control compound A, the inhibitory effect on the amplitude of pulsation was time-dependent, and it also had a significant effect on pulsatile frequency and regular index of pulsatile frequency. There was no significant effect on mitochondrial membrane potential. Conclusion: a feasible model for evaluating cardiac toxicity in vitro can be established by using human embryonic stem cell differentiation cardiomyocytes. The combination of real-time cell analysis and high intension cell imaging can be rapid and sensitive. Accurately evaluate the cardiac toxicity of the drug.
【作者单位】：中国食品药品检定研究院国家药物安全评价监测中心;中国科学院生物物理研究所;康龙化成(北京)新药技术股份有限公司;
【基金】：北京市科技计划课题资助项目(Z141100000214001)
【分类号】：R96

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