纳米氧化铜诱导小鼠肺泡巨噬细胞铜死亡的实验研究  

Experimental study on copper death of mouse alveolar macrophages induced by nano-copper oxide

作　　者：张晓晶 胡林霞 卜迁 孙东雷 ZHANG Xiao-jing;HU Lin-xia;BU Qian;SUN Dong-lei(Key Laboratory of Molecular Toxicology,Department of Education,West China School of Public Health,Sichuan University/West China Fourth Hospital,Chengdu,Sichuan 610041,China)

机构地区：[1]四川大学华西公共卫生学院/华西第四医院卫生毒理与病理学系,分子毒理四川省教育厅重点实验室,四川成都610041

出　　处：《现代预防医学》2024年第11期2066-2071,共6页Modern Preventive Medicine

基　　金：国家自然科学基金青年科学基金项目(82304110);四川大学专职博士后研发基金项目“粉尘颗粒诱导肺泡巨噬细胞铜死亡的作用及机制研究”。

摘　　要：目的探究纳米氧化铜诱导小鼠肺泡巨噬细胞铜死亡的毒性效应。方法利用MTT实验检测纳米氧化铜处理小鼠肺泡巨噬细胞(MH-S)存活率;透射电镜观察MH-S细胞形态;Western Blot检测铜死亡蛋白FDX1与DLAT的表达水平;转录组测序探究纳米氧化铜诱导铜死亡过程中细胞基因表达变化与关键信号通路;单因素方差分析进行差异分析。结果40 nm粒径的纳米氧化铜颗粒可以进入MH-S细胞内,引起MH-S细胞剂量-反应式的存活率下降(P<0.001),增加MH-S细胞内Cu^(2+)浓度,诱导FDX1(P=0.004)与DLAT蛋白(P<0.001)表达水平上升;而施加铜死亡抑制剂四硫钼酸盐(TTM)干预后减轻了细胞毒性,降低了细胞内Cu^(2+)水平(P=0.038),减少了铜死亡关键蛋白FDX1(P=0.012)与DLAT(P<0.001)表达。测序结果表明纳米氧化铜具有激活IL-17炎症通路的作用,而施加TTM干预后抑制了IL-17信号通路。结论纳米氧化铜具有诱导小鼠MH-S细胞铜死亡的毒性效应,为纳米氧化铜的毒作用机制研究提供了新线索。Objective To investigate the toxic effect of nano-copper oxide on copper death in mouse alveolar macrophages.Methods MTT assay was used to detect the survival rate of mouse alveolar macrophages(MH-S)treated with nano-copper oxide,transmission electron microscope to observe the morphology of MH-S cells,Western Blot to detect the expression of copper death protein FDX1 and DLAT,transcriptional group sequencing to explore the changes of cell gene expression and key signal pathways in the process of copper death induced by nano-copper oxide,and one-way ANOVA to analyze the difference.Results Copper oxide nanoparticles with 40nm size could enter MH-S cells,decrease the dose-response survival rate of MH-S cells(P<0.001),increase the concentration of Cu^(2+)in MH-S cells,and induce the expression of FDX1(P=0.004)and DLAT protein(P<0.001).However,the administration of copper death inhibitor tetra thiomolybdate(TTM)reduced the cytotoxicity,decreased the level of intracellular Cu^(2+)(P=0.0038),and decreased the expression of FDX1(P=0.012)and DLAT(P<0.05).The sequencing results showed that nano-copper oxide could activate IL-17 inflammatory pathway,while TTM could inhibit IL-17 signal pathway.Conclusion Nano-copper oxide can induce the death of copper in mouse MH-S cells,which provides a new clue for the study of the toxic mechanism of nano-copper oxide.

关 键 词：纳米氧化铜 肺泡巨噬细胞 铜死亡 

分 类 号：R114[医药卫生—卫生毒理学]