亚硫酸钠对人正常二倍体肝细胞的损伤作用程序性坏死相关蛋白受体相互作用蛋白1表达的影响  

Effects of sodium sulfite on damage and expression of necroptosis- associated protein-receptor interacting protein 1(RIP1) in HL-7702 cells

作　　者：杨舒筠 白剑英[1] 张艳[1] 王盼[1] 刘艳飞[1] 

机构地区：[1]山西医科大学公共卫生学院环境卫生学教研室,山西太原030001

出　　处：《环境与健康杂志》2015年第12期1061-1064,共4页Journal of Environment and Health

基　　金：山西省回国留学人员科研资助项目(2014-034);山西省留学回国人员科技活动择优资助项目(晋人社厅函[2014]376号)

摘　　要：目的探讨亚硫酸钠对人正常二倍体肝细胞(HL-7702)的损伤作用及程序性坏死相关蛋白受体相互作用蛋白1(RIP1)表达的影响。方法将处于对数生长期的HL-7702细胞分别暴露于含终浓度为0(对照)、0.01、0.1、1、10 mmol/L亚硫酸钠的培养基中培养2、4、12、24、48 h。采用CCK-8法检测细胞活性,采用全自动生化分析仪检测细胞上清液中丙氨酸氨基转移酶(ALT)、天冬氨酸氨基转移酶(AST)以及乳酸脱氢酶(LDH)的活力,采用Western Blot法检测细胞RIP1蛋白的表达水平。结果与对照组相比,1、10 mmol/L亚硫酸钠染毒不同时间的HL-7702细胞存活率降低,差异有统计学意义(P<0.05);且随着亚硫酸钠染毒浓度的升高,HL-7702细胞的存活率呈下降趋势。与对照组相比,仅10 mmol/L亚硫酸钠染毒24、48 h HL-7702细胞上清液中的ALT活力较高;1 mmol/L亚硫酸钠染毒4、24 h及10 mmol/L亚硫酸钠染毒2、4 h HL-7702细胞上清液中的AST活力较低,而10 mmol/L亚硫酸钠染毒24、48 h HL-7702细胞上清液中的AST活力较高;10 mmol/L亚硫酸钠染毒2、4 h HL-7702细胞上清液中的LDH活力较低,而染毒24、48 h HL-7702细胞上清液中的LDH活力较高,差异均有统计学意义(P<0.05)。与对照组相比,各浓度亚硫酸钠染毒组HL-7702细胞中RIP1蛋白的表达水平均较高,差异多有统计学意义(P<0.05)。结论程序性坏死可能参与了亚硫酸钠引起的肝细胞死亡,但其可能机制还有待于进一步研究。Objective To investigate the effects of sodium sulfite（Na_2SO_3） on damage and expression of necroptosis-associated proteinreceptor interacting protein 1（RIP1） in HL-7702 cells. Methods HL-7702 cells were exposed to the sodium sulfite medium with the final concentrations of 0, 0.01, 0.1, 1 and 10 mmol/L respectively, then cultured for 2, 4, 12, 24 and 48 h respectively.The cell viability was assayed by CCK-8 kit,the ALT,AST and LDH activities were detected by automatic biochemical analyzer,and the protein expression of RIP1 was tested by Western Blot. Results Compared with the control group,the survival rates of HL-7702 cells in 1 and 10 mmol/L Na_2SO_3-treated groups for 2-48 h were significantly decreased（P〈0.05）. With the increase of sodium sulfite dose,the survival rate of HL-7702 cells showed a decreasing trend. Compared with the control group,the activities of ALT were significantly increased in 10 mmol/L Na_2SO_3-treated groups for 24 h and 48 h（P〈0.05）. The activities of AST were significantly decreased in 1 mmol/L Na_2SO_3-treated groups for 2 h and 24 h and in 10 mmol/L Na_2SO_3-treated groups for 2 h and 4 h,but these were significantly increased in 10 mmol/L Na_2SO_3-treated groups for 24 h and 48 h（P 〈0.05）. The activities of LDH were significantly decreased in 10 mmol/L Na_2SO_3 treated groups for 2 h and 4 h, but these were significantly increased in 10 mmol/L Na_2SO_3-treated groups for 24 h and 48 h（P〈0.05）. Compared with the control group,the protein expression of RIP1 in all Na_2SO_3-treated groups were increased（P〈 0.05）. Conclusion The necroptosis may be involved in the process of sodium sulfite-induced hepatocyte death,but the possible mechanisms need further study.

关 键 词：亚硫酸钠 肝损伤 RIP1 细胞活性 

分 类 号：R994.6[医药卫生—毒理学]