亚砷酸钠对小鼠肝细胞AML12氧化应激、凋亡及MST1、MST2蛋白表达的影响  

Effects of sodium arsenite on oxidative stress,apoptosis,and the expression of MST1,MST2 in mouse hepatocytes AML12

作　　者：赵哲仪 王正蓉 方兴艳 王甜 罗昭逊[5] 谢婷婷 ZHAO Zheyi;WANG Zhengrong;FANG Xingyan;WANG Tian;LUO Zhaoxun;XIE Tingting(Clinical Laboratory Center,the Affiliated Hospital of Guizhou Medical University,Guiyang 550004,Guizhou,China;School of Clinical Laboratory Science,Guizhou Medical University,Guiyang 550004,Guizhou,China;Prenatal Diagnosis Center,the Affiliated Hospital of Guizhou Medical University,Guiyang 550004,Guizhou,China;Department of Clinical Laboratory,Shandong Provincial Third Hospital,Jinan 250031,Shandong,China;Academy of Pediatrics,Guizhou Medical University,Guiyang 550004,Guizhou,China)

机构地区：[1]贵州医科大学附属医院临床检验中心,贵州贵阳550004 [2]贵州医科大学医学检验学院,贵州贵阳550004 [3]贵州医科大学附属医院产前诊断中心,贵州贵阳550004 [4]山东省立第三医院检验科,山东济南250031 [5]贵州医科大学儿科学院,贵州贵阳550004

出　　处：《贵州医科大学学报》2022年第1期7-12,共6页Journal of Guizhou Medical University

基　　金：国家自然科学基金(81560514);贵州省科技创新人才团队项目(2019-5610)。

摘　　要：目的探讨亚砷酸钠(NaAsO_(2))对小鼠肝细胞AML12氧化应激、凋亡及哺乳动物不育系20样激酶1(MST1)、MST2蛋白表达的影响。方法取对数生长期的小鼠正常肝细胞AML12,分为对照组(不含NaAsO_(2))及实验组(10、15、20、25及30μmol/L NaAsO_(2)处理24 h),采用荧光探针染色法和化学比色法分别检测各组小鼠AML12细胞内活性氧(ROS)含量和超氧化物歧化(SOD)活性水平,采用Western blot检测各组小鼠AML12细胞的MST1、MST2、磷酸化MST1/2(p-MST1/2)及半胱氨酸蛋白酶3剪接体(c-caspase 3)蛋白相对表达量。结果与对照组相比,随着NaAsO_(2)浓度的增加,各实验组小鼠AML12细胞内ROS含量逐渐升高(P<0.05),SOD活性逐渐降低(P<0.05);与对照组相比,各实验组小鼠AML12细胞内p-MST1/2蛋白相对表达量升高(P<0.05),20-30μmol/L NaAsO_(2)组小鼠AML12细胞内为MST1蛋白相对表达量降低、c-caspase 3蛋白相对表达量升高(P<0.05),15-30μmol/L NaAsO_(2)组小鼠AML12细胞内MST1蛋白相对表达量升高(P<0.05)。结论NaAsO_(2)可促进小鼠肝细胞AML12发生氧化应激、凋亡及p-MST1/2、MST1、MST2蛋白表达异常。Objective To investigate the effects of sodium arsenite(NaAsO_(2))on oxidative stress,apoptosis and expression of mammalian sterile 20-like kinases 1(MST1),MST2 in mouse normal hepatocytes AML12.Methods Mouse normal hepatocytes AML12 at logarithmic growth stage were divided into control group and experimental group.Cells in control group were not treated with NaAsO_(2).Cells in experimental group were treated with 10,15,20,25,and 30μmol/L NaAsO_(2) for 24 h.The content of intracellular reactive oxygen species(ROS)and the activity of superoxide dismutase(SOD)were detected by fluorescent probe technique and colorimetry respectively.The relative expression levels of p-MST1/2,MST1,MST2,and cleaved caspase3(c-caspase 3)protein were detected by Western blot.Results Compared with the control group,the ROS content of AML12 cells increased gradually(P<0.05),while the activity of SOD decreased gradually along with the increase of NaAsO_(2) concentration(P<0.05);Compared with the control group,the relative expression of p-MST1/2 protein in NaAsO_(2) groups increased gradually(P<0.05),the relative expression of MST1 protein in 20-30μmol/L NaAsO_(2) groups decreased(P<0.05),the relative expression of c-caspase 3 in 20-30μmol/L NaAsO_(2) groups and the relative expression of MST2 in 15-30μmol/L NaAsO_(2) groups increased(P<0.05).Conclusion NaAsO_(2) may induce oxidative stress,apoptosis,and the abnormal expression of p-MST1/2,MST1 and MST2 in mouse normal hepatocytes AML12.

关 键 词：细胞凋亡 亚砷酸钠 肝细胞AML12 氧化应激 哺乳动物不育系20样激酶1 哺乳动物不育系20样激酶2 

分 类 号：R994.6[医药卫生—毒理学]