摘要
目的探讨纳米氧化锌(Zinc Oxide Nanoparticles,ZnO NPs)对人源心肌细胞(A human cardiomyocytes,AC16)的毒性作用特征,并从线粒体自噬角度探讨其作用机制。方法利用透射电镜(Transmission electron microscope,TEM)和动态光散射法(Dynamic light scattering,DLS)对ZnO NPs进行表征,将AC16细胞给予不同剂量的ZnO NPs处理,光学显微镜观察细胞形态学改变,利用CCK-8法测定细胞存活率。流式细胞术检测线粒体活性氧自由基(Reactive oxygen species,ROS)和线粒体膜电位(Mitochondrial membrane potential,MMP)改变,Western blot检测线粒体自噬蛋白PINK1和Parkin的表达水平。结果透射电镜显示ZnO NPs为棒状结构,平均粒径约为50 nm,Zeta电位为-17.57±3.07 mV。ZnO NPs呈剂量-时间依赖性降低AC16细胞存活率,导致线粒体功能损伤,表现为线粒体ROS蓄积和MMP下降。细胞给予50μmol/L ZnO NPs处理6 h可增加PINK1和Parkin的表达,提示线粒体自噬激活;线粒体自噬抑制剂(Mitochondrial division inhibitor,Mdivi-1)可抑制PINK1和Parkin蛋白表达,加剧ZnO NPs引起的细胞毒性。结论ZnO NPs可剂量和时间依赖性引起人源心肌细胞毒性和线粒体功能紊乱,提示线粒体自噬可作为一种细胞自我保护机制而抑制ZnO NPs的毒性。
Objective To investigate the toxic effects of zinc oxide nanoparticles(ZnO NPs)on human cardiomyocytes(AC16)and to explore the mechanism in terms of mitophagy regulation.Methods The morphological characteristic of ZnO NPs was assayed by transmission electron microscope(TEM)and dynamic light scattering(DLS).After the AC16 cells were treated with different concentrations of ZnO NPs,the cell morphology was observed under light microscope.The cell viability was measured by CCK-8 method.The reactive oxygen species(ROS)and mitochondrial membrane potential(MMP)were detected by flow cytometry method,and the protein expressions of PINK1 and Parkin were detected by Western blot.Results TEM showed that ZnO NPs were rod-like structure with an average particle size of about 50 nm.Zeta potential of ZnO NPs was-17.57±3.07 mV.ZnO NPs decreased the cell viability in a concentration-and time-dependent manner,and induced mitochondrial dysfunction as evidenced by ROS accumulation and MMP loss.After the AC16 cells were treated with 50μmol/L ZnO NPs for 6 h,the protein expressions of PINK1 and Parkin were increased,indicating the activation of mitophagy.A mitophagy inhibitor(Mdivi-1)was found to decrease the protein expressions of PINK1 and Parkin and to enhance the cytotoxicity induced by ZnO NPs.Conclusion It was demonstrated that ZnO NPs concentration-and time-dependently elicited toxicity and mitochondrial dysfunction in human cardiomyocytes,and suggested that mitophagy exert self-protective role to inhibit the toxicity of ZnO NPs.
作者
张林聪
李宇杰
尹圆圆
张弛
彭辉
郭家彬
武冬梅
ZHANG Lin-cong;LI Yu-jie;YIN Yuan-yuan;ZHANG Chi;PENG Hui;GUO Jia-bin;WU Dong-mei(Jiamusi University,jiamusi Heilongjiang 154007,China;Center for Disease Control and Prevention,PLA,Beijing 10071,China)
出处
《毒理学杂志》
CAS
CSCD
2021年第2期117-122,共6页
Journal of Toxicology
基金
国家重研发计划课题(2018YFC1602602)
北京市科技新星项目(Z171100001117103)
食品安全毒理学研究与评价北京市重点实验室开放基金(KF2019-01)。
关键词
纳米氧化锌
心肌细胞
线粒体
自噬
Zinc oxide nanoparticles
Cardiomyocytes
Mitochondria
Autophagy
