摘要
目的观察亚砷酸钠(NaAsO2)对人正常肝细胞(L-02细胞)脂代谢基因固醇调节元件结合蛋白-1c(SREBP-1c)、过氧化物酶体增殖物激活受体α(PPARα)和脂肪酸合成酶(FAS)表达的影响。方法体外培养L-02细胞,分别以0(对照)、2、4、8、16、32、64、128μmol/L NaAsO2染砷24 h,采用CCK-8法检测细胞存活率,并制作拟合曲线计算半抑制浓度(IC50),以IC50的0、1/8、1/4、1/2作为染砷剂量进行后续实验。采用甘油磷酸氧化酶-过氧化氢酶(GPO-PAP)法检测细胞甘油三酯(TG)含量;实时荧光定量PCR检测SREBP-1c、PPARα、FAS mRNA表达水平;蛋白质印迹法(Western blot)检测SREBP-1c和PPARα蛋白表达水平。结果8、16、32、64、128μmol/L NaAsO2组细胞存活率[(92.000±1.414)%、(91.000±0.000)%、(76.500±0.707)%、(53.000±1.412)%、(47.000±1.412)%]明显低于对照组[(100.000±0.000)%,P均<0.01],IC50为64μmol/L,以0(对照)、8、16、32μmol/L NaAsO2进行后续实验。与对照组[(1.000±0.000)mmol/g prot]比较,8、16、32μmol/L NaAsO2组TG含量[(0.691±0.064)、(0.474±0.162)、(0.184±0.045)mmol/g prot]显著降低(P均<0.01)。与对照组比较,各染砷组SREBP-1c、PPARα、FAS mRNA表达水平,SREBP-1c、PPARα蛋白表达水平显著降低(P<0.01或<0.05)。相关性分析可见,NaAsO2含量与细胞TG含量,SREBP-1c和PPARα蛋白表达水平呈负相关(r=-0.954、-0.875、-0.965,P均<0.01)。结论NaAsO2可引起L-02细胞TG含量减少及脂代谢相关基因SREBP-1c、PPARα和FAS表达降低,提示砷致肝损伤过程中可引起脂代谢障碍发生。
Objective To investigate the effects of sodium arsenite(NaAsO2)on the expression of sterol regulatory element-binding protein-1c(SREBP-1c),peroxisome proliferator activated receptorα(PPARα)and fatty acid synthase(FAS)in human liver cells(L-02 cells).Methods L-02 cells were cultured in vitro,and exposed to NaAsO2 at 0(control),2,4,8,16,32,64 and 128μmol/L for 24 h,respectively,and the cell survival rate was determined by CCK-8 method.And a fitting curve was made to calculate the half inhibitory concentration(IC50),subsequent experiments were carried out with 0,1/8,1/4 and 1/2 of IC50 as arsenic exposure doses.Glycerol phosphate oxidase-catalase(GPO-PAP)method was used to detect the content of triglyceride(TG)in cells;the mRNA expression levels of SREBP-1c,PPARαand FAS were detected by Real-time PCR;and the protein expression levels of SREBP-1c and PPARαwere detected by Western blotting.Results The cell survival rates of 8,16,32,64 and 128μmol/L NaAsO2 groups[(92.000±1.414)%,(91.000±0.000)%,(76.500±0.707)%,(53.000±1.412)%,(47.000±1.412)%]were significantly lower than that of the control group[(100.000±0.000)%,P<0.01].The IC50 was 64μmol/L,and subsequent experiments were conducted with 0(control),8,16 and 32μmol/L NaAsO2,respectively.Compared with the control group[(1.000±0.000)mmol/g prot],TG contents of 8,16 and 32μmol/L NaAsO2 groups[(0.691±0.064),(0.474±0.162),(0.184±0.045)mmol/g prot]were significant decreased(P<0.01).Compared with the control group,the mRNA expression levels of SREBP-1c,PPARα,FAS,and the protein expression levels of SREBP-1c and PPARαin NaAsO2 groups were significantly decreased(P<0.01 or<0.05).Correlation analysis showed that NaAsO2 content was negatively correlated with TG content,SREBP-1c and PPARαprotein expression levels(r=-0.954,-0.875,-0.965,P<0.01).Conclusion NaAsO2 can reduce the TG content and the expression of lipid metabolism related genes SREBP-1c,PPARαand FAS in L-02 cells,suggesting that arsenic-induced liver injury can cause lipid metabolism disorders.
作者
张琦
谢万生
段恬筱
李晓之
简文
吴长艳
胡婷
罗鹏
Zhang Qi;Xie Wansheng;Duan Tianxiao;Li Xiaozhi;Jian Wen;Wu Changyan;Hu Ting;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254-258,共5页
Chinese Journal of Endemiology
基金
国家自然科学基金(81660835、81860560、U1812403)
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)
贵州省教育厅青年科技人才成长项目(黔教合KY字[2018]188号)。
