线粒体膜通透性在微囊藻毒素诱导的小鼠肝细胞凋亡中的作用  

Effect of membrane permeability transition on hepatocyte apoptosis of the microcystin-LR-induced mice

作　　者：刘菁[1] 魏慇娜 沈萍萍[1] 

机构地区：[1]南京大学医药生物技术国家重点实验室,南京210093

出　　处：《卫生研究》2011年第1期53-56,共4页Journal of Hygiene Research

基　　金：国家重点基础研究发展计划项目(No.2008CB418102);江苏省自然科学基金重点项目(No.BK2010049)

摘　　要：目的从线粒体膜通透性(MPT)的角度去研究微囊藻毒素(MCs)诱导小鼠肝细胞凋亡的作用机制,并揭示活性氧(ROS)在其中的作用。方法将ICR雄性小鼠随机分成4组,A组:腹腔注射生理盐水;B组:腹腔注射MC-LR 50μg/kg bw;C组:腹腔注射线粒体膜通透性抑制剂环孢霉素A(CsA)100mg/kg bw;D组:提前1小时腹腔注射CsA 100mg/kg bw后再腹腔注射MC-LR 50μg/kg bw。分离小鼠血清,检测AST、ALT含量;分离小鼠肝脏,进行小鼠肝脏DNA片段化检测、线粒体膜电位检测、ROS水平检测和Bcl-2表达水平的western blotting检测。结果在50μg/kg bw MC-LR处理组中,血清中AST、ALT水平升高,肝细胞中出现明显的DNA ladder、Bcl-2蛋白水平的表达上调、肝组织线粒体膜电位降低、组织ROS水平升高;100mg/kgbw CsA处理组小鼠各项指标没有明显变化;而预处理100mg/kg bw CsA后再腹腔注射50μg/kg bw MC-LR则可以消除肝组织DNA ladder,并且使血清AST、ALT含量,肝脏Bcl-2蛋白的表达、线粒体膜电位和ROS水平恢复到对照组的水平。结论体内环境下,抑制线粒体膜转运通道可抑制MC-LR诱导的肝细胞凋亡。Objective From the view of membrane permeability transition（MPT） to study the mechanism of microcystin（MCs） inducing hepatocyte apoptosis in mice,and unclose the function of reactive oxygen species（ROS） in this process.Methods Use the mice as the model animals and randomly divide them into 4 groups.Group A were injected with physiological saline peritoneal in abdomen,while group B were injected with MC-LR 50μg /kg bw in abdomen,group C were injected with the MPT inhibitor,Cyclosporin A（CsA） 100mg /kg bw,in abdomen,and group D were injected with CsA 100mg /kg bw in abdomen,and then after 1 hour,with MC-LR 50μg /kg bw.Separate the mice serum,measure AST and ALT concentrations.Separate the mice livers and then perform the mice liver DNA segment detection,mitochondrion membrane potential level measurement,ROS level measurement and western blotting measurement at the expression level of Bcl-2.Results In the group treated with 50μg /kg bw MC-LR,in the serum,the AST and ALT levels rised.In the liver cells,there was significant DNA ladder,the the expression of Bcl-2 at the level of protein increased,mitochondrion membrane potential level of liver tissue decreased,and the ROS level in the tissue increased.In the group treated with 100mg /kg bw CsA,every index of the mice has no significant change.While with the pre-treatment of 100mg /kg bw CsA,and then with 50μg /kg bw MC-LR in abdomen,the DNA ladder in the liver tissue can be removed,and the serum AST and ALT concentrations,the expression of liver Bcl-2 protein,the mitochondrion membrance potentiial level and ROS level were conversed to the levels of the control group.Conclusion In vivo the inhibition of transition chanel of mitochondrion membrance can inhibit the hepatocyte apoptosis induced by MC-LR.

关 键 词：微囊藻毒素 细胞凋亡 线粒体膜通透性 肝细胞 

分 类 号：R994.6[医药卫生—毒理学] Q593.1[医药卫生—药学]