亚砷酸钠对人正常肝细胞凋亡及Bax和Bcl-2 mRNA和蛋白表达的影响  

Effect of sodium arsenite on human hepatocytes apoptosis and expression of Bax,Bcl-2 mRNA and protein

作　　者：张豫[1] 罗鹏[1] 叶建方[1] 肖佳艳[1] 吴雪艳[1] 丁锴[1] 张吉梅[1] 

机构地区：[1]贵阳医学院公共卫生学院卫生毒理教研室,贵州贵阳550004

出　　处：《环境与健康杂志》2014年第12期1048-1051,共4页Journal of Environment and Health

基　　金：国家自然科学基金(81160336);贵州省科技厅农业攻关计划项目(黔科合NY字[2011]3055号);贵州省教育厅基金项目(黔省专合字[2010]39号);贵州省科技厅(黔科合J字[2010]2188)

摘　　要：目的探讨亚砷酸钠对人正常肝(L-02)细胞凋亡及Bax、Bcl-2 m RNA和蛋白表达的影响。方法将L-02肝细胞分别暴露于含0(对照)、50、100、150μmol/L亚砷酸钠的培养基中暴露24 h。采用MTT法检测L-02肝细胞的存活率,采用流式细胞术检测L-02肝细胞凋亡率,采用RT-PCR的方法检测Bax、Bcl-2 m RNA相对表达情况,采用Westernblotting方法检测L-02肝细胞中Bax、Bcl-2蛋白的表达情况。结果与对照组相比,各浓度亚砷酸钠染毒组L-02细胞的存活率均较低,凋亡率均较高,差异均有统计学意义(P<0.05);且随着亚砷酸钠染毒浓度的升高,L-02细胞的存活率呈下降趋势,而凋亡率上升趋势。与对照组相比,各浓度亚砷酸钠染毒组L-02细胞中Bcl-2、Bax m RNA和蛋白的表达水平均较高,100μmol/L亚砷酸钠染毒组Bcl-2 m RNA/Bax m RNA值也较高,而150μmol/L亚砷酸钠染毒组Bcl-2 m RNA/Bax m RNA值及50、150μmol/L亚砷酸钠染毒组Bcl-2蛋白/Bax蛋白值均较低,差异均有统计学意义(P<0.05)。结论亚砷酸钠可抑制L-02肝细胞的生长,诱导细胞凋亡的发生;其机制可能与Bcl-2家族的激活有关。Objective To investigate the impact of different doses of sodium arsenite（NaAsO2） on L-02 hepatocytes apoptosis and the expression of Bcl-2,Bax mRNA and protein. Methods The L-02 cells were treated with Na As O2 at the doses of0,50,100 and 150 μmol/L respectively for 24 h. MTT was used to detect cell viability. The ratios of expressions of Bax and Bcl-2 mRNA were detected by RT-PCR. Western-blotting was used to investigate the protein levels of total Bcl-2 and Bax.Results Compared with the control,in arsenic-exposed groups the survival rates were significant lower and the apoptosis rates were significant higher（P〈0.05） with dose-dependent manner,and the expression of Bcl-2 and Bax m RNA and total protein levels were higher,the Bcl-2 mRNA/Bax m RNA value was higher in 100 μmol/L Na As O2 exposure group,the Bcl-2 mRNA/Bax m RNA value in 150 μmol/L NaAsO2 exposure group and the Bcl-2 protein/Bax protein value in 50,150 μmol/L NaAsO2 exposure groups were significant lower（P〈0.05）. Conclusion Sodium arsenite can inhibit the growth of L-02 liver cells and induce apoptosis; The mechanism may be related to the activation of the Bcl-2 family.

关 键 词：亚砷酸钠 细胞凋亡 基因 BAX BCL-2 

分 类 号：R994.6[医药卫生—毒理学]