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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 To study the mechanism of microcystin (MCs) -induced hepatocyte apoptosis in mitochondrial membrane permeability (MPT) and to reveal the role of reactive oxygen species (ROS) in it. Methods ICR male mice were randomly divided into 4 groups: group A: intraperitoneal injection of saline; group B: intraperitoneal injection of MC-LR 50 μg / kg bw; group C: intraperitoneal injection of mitochondrial membrane permeability inhibitor cyclosporine A CsA) 100mg / kg bw; Group D: MC-LR 50μg / kg bw was intraperitoneally injected CsA 100mg / kg bw one hour earlier. The serum of mice was separated and the contents of AST and ALT were detected. The liver of mice was isolated and the hepatic DNA fragmentation, mitochondrial membrane potential, ROS level and Bcl-2 expression were detected by western blotting. Results In the 50μg / kg bw MC-LR treatment group, the levels of AST and ALT in serum increased, and DNA ladder appeared in hepatocytes, the expression of Bcl-2 protein was up-regulated, the mitochondrial membrane potential of liver tissue was decreased, And 100 mg / kg bw CsA had no significant changes in all the indexes. However, pretreatment with 100 mg / kg bw CsA followed by intraperitoneal injection of 50 μg / kg bw MC-LR could abolish the DNA ladder in liver tissue and serum AST, ALT levels, liver Bcl-2 protein expression, mitochondrial membrane potential and ROS levels returned to the control group. Conclusion In vivo, inhibition of mitochondrial membrane transport channels can inhibit MC-LR-induced hepatocyte apoptosis.