摘要
目的探讨核因子-E2相关因子2(Nrf2)信号通路在亚砷酸钠(NaAsO2)致人正常肝细胞(L-02细胞)氧化损伤过程中的作用,为砷致肝损伤的氧化损伤作用机制研究提供实验依据。方法体外培养L-02细胞,分别以0(对照)、25、50、75、100、125、150μmol/L NaAsO2处理细胞24 h,采用CCK8法检测细胞存活率;根据细胞存活率计算半抑制浓度(IC50),分别以IC50的0、1/8、1/4、1/2倍剂量NaAsO2处理L-02细胞进行分组实验。采用蛋白免疫印迹法(Western blot)检测L-02细胞中和细胞核内Nrf2信号通路相关因子Nrf2、血红素氧化酶-1(HO-1)、醌氧化还原酶1(NQO1)、谷胱甘肽过氧化物酶1(GPx1)的蛋白表达情况。结果CCK8实验结果显示,25、50、75、100、125、150μmol/L NaAsO2组的L-02细胞存活率[(69.53±0.06)%、(41.33±0.08)%、(23.65±0.04)%、(26.51±0.04)%、(31.63±0.01)%、(26.24±0.02)%]明显低于对照组[(100.00±0.00)%,P均<0.05];细胞存活率的IC50为40μmol/L,分组实验的NaAsO2剂量分别采用0(对照)、5、10、20μmol/L。Western blot检测结果显示,与对照组比较,5、10、20μmol/L NaAsO2组L-02细胞中Nrf2、HO-1及L-02细胞核内HO-1蛋白水平显著升高(P均<0.05);10、20μmol/L NaAsO2组L-02细胞中GPx1蛋白水平显著降低(P均<0.05),L-02细胞核内Nrf2蛋白水平显著升高(P均<0.05);5μmol/L NaAsO2组L-02细胞核内NQO1蛋白水平显著升高(P<0.05)。结论NaAsO2对L-02细胞中Nrf2信号通路相关因子的表达有影响,其致L-02细胞氧化损伤的作用机制可能与Nrf2信号通路有关。
Objective To explore the role of nuclear factor-E2-related factor 2(Nrf2)signaling pathway in oxidative damage caused by sodium arsenite(NaAsO2)in human normal liver cells(L-02),and to provide experimental basis for the study of oxidative damage mechanism of liver damage caused by arsenic.Methods L-02 cells were cultured in vitro and treated with 0(control),25,50,75,100,125,and 150μmol/L NaAsO2,respectively,for 24 h.The half-inhibitory concentration(IC50)was calculated according to the cell survival rate by CCK8,and L-02 cells were treated with 0,1/8,1/4 and 1/2 dose of IC50 of NaAsO2,respectively,for grouping experiments.Protein expressions of Nrf2,heme oxygenase-1(HO-1),NADH quinone oxidoreductase 1(NQO1)and glutathione peroxidase 1(GPx1)in L-02 cells and L-02 nucleus were detected by Western blotting.Results The result of CCK8 showed that the survival rates of L-02 cells in 25,50,75,100,125,150μmol/L NaAsO2 groups were[(69.53±0.06)%,(41.33±0.08)%,(23.65±0.04)%,(26.51±0.04)%,(31.63±0.01)%,(26.24±0.02)%],which were significantly lower than that of the control group[(100±0.00)%].The differences were statistically significant(P<0.05).The IC50 calculated by cell survival was 40μmol/L,and the NaAsO2 doses used in the experiment were 0(control),5,10,and 20μmol/L.Western blotting results showed that,compared with the control group,the protein expression levels of Nrf2,HO-1 in L-02 and HO-1 in the L-02 cells nucleus in the 5,10 and 20μmol/L NaAsO2 groups were significantly higher(P<0.05).Compared with the control group,the expression levels of GPx1 protein in L-02 cells of 10 and 20μmol/L NaAsO2 groups were decreased(P<0.05).Compared with the control group,the expression levels of Nrf2 protein in L-02 nucleus in 10 and 20μmol/L NaAsO2 groups were significantly increased(P<0.05);the expression level of NQO1 protein in L-02 nucleus in 5μmol/L NaAsO2 group was significantly increased(P<0.05).Conclusion NaAsO2 has an effect on the expression of Nrf2 signaling pathway related factors in L-02 cells,and the mechanism of oxidative damage caused by NaAsO2 in L-02 cells may be related to Nrf2 signaling pathway.
作者
李晓之
胡婷
段恬筱
张琦
吴长艳
简文
罗鹏
Li Xiaozhi;Hu Ting;Duan Tianxiao;Zhang Qi;Wu Changyan;Jian Wen;Luo Peng(School of Public Health,Guizhou Medical University,Guiyang 550025,China;Key Laboratory for Environmental Pollution Monitoring and Disease Control,Ministry of Education,Guizhou Medical University,Guiyang 550025,China)
出处
《中华地方病学杂志》
CAS
CSCD
北大核心
2020年第4期259-263,共5页
Chinese Journal of Endemiology
基金
国家自然科学基金(81660835、81860560、U1812403)
贵州省教育厅青年科技人才成长项目(黔教合KY字[2018]188号)
贵州省区域内一流学科建设项目-公共卫生与预防医学(黔教科研发[2017]85号)。
