铜通过活性氧诱导鸡肝细胞发生焦亡  

Copper induces pyroptosis by generating ROS in chicken hepatocytes

作　　者：廖建昭 杨帆[1] 裴若男 刘德琴 陈慧莲 张卓炜 张裕恒 唐兆新[1] LIAO Jian-zhao;YANG Fan;PEI Ruo-nan;LIU De-qin;CHEN Hui-lian;ZHANG Zhuo-wei;ZHANG Yu-heng;TANG Zhao-xin(College of Veterinary Medicine,South China Agricultural University,Guangzhou 510642,China)

机构地区：[1]华南农业大学兽医学院

出　　处：《中国兽医学报》2018年第12期2374-2378,共5页Chinese Journal of Veterinary Science

基　　金：国家自然科学基金资助项目(31572585)

摘　　要：为探讨铜是否能诱导肝细胞发生焦亡及其发生机制,本研究通过培养鸡原代肝细胞,以不同浓度的硫酸铜(0,10,50,100μmol/L)处理细胞24h,CCK-8法检测细胞活力,实时荧光定量PCR检测细胞焦亡相关基因Caspase-1、IL-1β、IL-18、NLRP3的mRNA转录水平,酶联免疫吸附法(ELISA)测定细胞上清液中焦亡相关蛋白Caspase-1、IL-1β、IL-18的含量,并使用活性氧清除剂N-乙酰半胱氨酸(NAC)与铜共同作用于肝细胞后观察上述指标的变化。结果显示,铜可诱导细胞存活率下降(P<0.05),焦亡相关基因Caspase-1、IL-1β、IL-18和NLRP3表达显著上调(P<0.05),并促使细胞上清液中Caspase-1、IL-1β和IL-18的含量升高(P<0.05);NAC与铜联合作用可使肝细胞存活率较铜单独作用显著升高(P<0.05),焦亡相关基因表达显著下调(P<0.05),相关蛋白含量也显著降低(P<0.05)。结果表明,铜可通过活性氧诱导鸡肝细胞发生焦亡。To investigate whether copper(Cu)can induce pyroptosis in hepatocytes and its mechanism,primary chicken hepatocytes were treated with different concentrations of Cu sulfate(CuSO4)(0,10,50,and 100μmol/L)for 24 h.Cell viability was determined using CCK-8 assays.Additionally,the mRNA expression levels of pyroptosis-related genes(Caspase-1,IL-1β,IL-18,and NLRP3)were detected by real-time quantitative PCR.Moreover,the contents of pyroptosis-related proteins in the supernatant fluid of hepatocytes were also measured by enzyme-linked immunosorbent assay(ELISA).Furthermore,hepatocytes were treated with N-acetylcysteine(NAC)and Cu to determine above factors.Results showed that Cu can induce cell viability decreased(P<0.05).The mRNA expression levels of Caspase-1,IL-1β,IL-18,and NLRP3 in treatment groups significantly upregulated compared to control(P<0.05).Additionally,the contents of Caspase-1,IL-1β,and IL-18 in the supernatant fluid of hepatocytes were also remarkably increased(P<0.05).Moreover,cell viability was markedly increased,pyroptosis-related genes and proteins above were significantly decreased in cells treated with NAC and Cu compared to single Cu treatment(P<0.05).These results suggest that Cu can induce pyroptosis by generating reactive oxygen species(ROS)in chicken hepatocytes.

关 键 词：铜 焦亡 鸡 肝细胞 活性氧 

分 类 号：S856.9[农业科学—临床兽医学]