Involvement of oxidative stress in ZnO NPs-induced apoptosis and autophagy of mouse GC-1 spg cells
Involvement of oxidative stress in ZnO NPs-induced apoptosis and autophagy of mouse GC-1 spg cells
复制标题
氧化应激参与 ZnO NPs 诱导小鼠 GC-1 spg 细胞凋亡和自噬
DOI：
10.1016/j.ecoenv.2020.110960
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发表时间：
2020
影响因子：
6.8
通讯作者：
Jiaxiang Chen
中科院分区：
文献类型：
--
作者：
Dan Yang;Meijuan Zhang;Yu Gan;Si Yang;Jinglei Wang;Mei Yu;Jie Wei;Jiaxiang Chen
氧化锌纳米颗粒（ZnO NPs）已被广泛应用于各个行业，并被报道抑制精子发生，然而，ZnO NPs诱导的精子发生失败尚未完全阐明。本研究采用自由基清除剂N-乙酰-L-半胱氨酸（NAC）或自噬抑制剂3-甲基腺嘌呤（3-MA）分别处理小鼠精原细胞系GC-1 spg细胞24 h，MTT法检测细胞活力，Western blotting法检测细胞凋亡，AnnexinV-FITC/PI法检测细胞凋亡率，MTT法检测细胞凋亡率。采用免疫印迹法和透射电镜检测细胞自噬，氧化应激试剂盒检测MDA、GSH含量和SOD、GSH-PX活性。本研究表明，ZnO纳米颗粒暴露抑制小鼠GC-1 spg细胞的活力并诱导其凋亡。有趣的是，ZnO纳米颗粒显着增加LC 3-II的蛋白质含量，LC 3-II/LC 3-I的比例，以及ATG 5和Beclin 1的蛋白质水平在细胞中。此外，透射电子显微镜（TEM）显示，经ZnO NPs处理的细胞胞浆中的自噬囊泡显着增加，表明ZnO NPs可以诱导细胞的自噬。ZnO纳米颗粒可诱导细胞产生氧化应激，抑制氧化应激可减轻ZnO纳米颗粒诱导的细胞凋亡和自噬。3-MA对自噬的抑制可以缓解ZnO纳米颗粒对细胞活力的抑制和对细胞凋亡的诱导，表明自噬对ZnO纳米颗粒诱导的细胞凋亡具有细胞毒性作用。综上所述，氧化应激参与了ZnO NPs诱导的小鼠GC-1 spg细胞凋亡和自噬，自噬可能在ZnO NPs诱导的细胞凋亡中发挥细胞毒性作用。
Zinc oxide nanoparticles (ZnO NPs) have been extensively used in various industries and reported to inhibit spermatogenesis, however, ZnO NPs-induced spermatogenesis failure is yet to be fully elucidated. Herein, mouse-derived spermatogonia cell line GC-1 spg cells were treated with ZnO NPs for 24 h in the presence or absence of radical scavenger N-acetyl-L-cysteine (NAC) or autophagy inhibitor 3-methyladenine (3-MA), then cell viability was observed by MTT assay; apoptosis was observed by western blotting analysis and AnnexinV-FITC/PI assay, respectively; autophagy was detected by western blotting analysis and transmission electron microscopy, respectively; and the contents of MDA and GSH and the activities of SOD and GSH-PX were measured by oxidative stress kits. The present study showed that ZnO NPs exposure inhibited viability and induced apoptosis of mouse GC-1 spg cells. Intriguingly, ZnO NPs markedly increased the protein content of LC3-II, the ratio of LC3-II/LC3-I, and the protein levels of ATG 5 and Beclin 1 in the cells. Furthermore, transmission electron microscopy (TEM) showed that autophagic vesicles in the cytoplasm increased significantly in the ZnO NPs-treated cells, indicating that ZnO NPs could induce autophagy of the cells. Oxidative stress could be induced by ZnO NPs; moreover, inhibition of oxidative stress could alleviate the induction of apoptosis and autophagy by ZnO NPs. Inhibition of autophagy by 3-MA could rescue the inhibition of cell viability and induction of apoptosis by ZnO NPs, which indicated that autophagy might have cytotoxic effect on ZnO NPs-induced apoptosis. In summary, oxidative stress was involved in ZnO NPs-induced apoptosis and autophagy of mouse GC-1 spg cells, and autophagy might play a cytotoxic role in ZnO NPs-induced apoptosis.