紫菀肽类组分诱导肝脏L-02细胞凋亡的机制  

Mechanism of apoptosis induced by peptide-rich fraction from the root of Aster tataricus in liver L-02 cells

作　　者：王丽[1] 李明丹[1] 卢毅[1] 张朝凤[1] 许翔鸿[1] 张勉[1] 

机构地区：[1]中国药科大学生药学研究室,南京211198

出　　处：《中国药科大学学报》2014年第4期469-474,共6页Journal of China Pharmaceutical University

基　　金：国家自然科学基金资助项目(No.30772702);中国药科大学大学生创新创业训练计划(No.SZ13116)~~

摘　　要：探讨紫菀肽类组分Fr-2的肝细胞毒性及其作用机制。采用二甲基噻唑蓝(MTT)检测人正常肝脏L-02细胞的活力,试剂盒检测LDH、ROS、GSH、细胞色素C和Caspase-9、Caspase-3的含量,流式细胞仪检测细胞凋亡率和线粒体膜电位变化,Western blot法检测凋亡相关蛋白p-JNK、Bax、Bcl-2的表达。实验结果显示,紫菀肽类组分Fr-2可剂量/时间依赖性地抑制L-02细胞的生长,诱导细胞内的氧化应激反应,降低线粒体膜电位,促进细胞色素C的释放,升高Bax/Bcl-2的比值并激活Caspase-9、Caspase-3。以上结果表明,紫菀肽类组分Fr-2导致肝细胞毒性的主要诱因是氧化应激,主要作用机制是诱导线粒体依赖途径的细胞凋亡。To evaluate the hepatotoxicity and its mechanism of the peptide-rich fraction （Fr-2） of the root of Aster tataricus in human liver L-02 cells. Cell viability was assessed by MTT assay and the contents of LDH, ROS, GSH, cytoehrome C and Caspase-9/3 were assessed by commercially available kits. Cell apoptosis and mito- chondrial membrane potential were measured by flow cytometry. The protein expressions of p-JNK, Bax, Bcl-2 were analysed by Western blot assay. The results demonstrated that the Fr-2 fraction induced apoptosis in a concentration- and time-dependent manner and provoked oxidative stress-associated inflammation in L-02 cells. It was characterized by loss of mitochondrial membrane potential and release of cytochrome c from mitochondria. The enhanced Bax/Bcl-2 ratio and the activated Caspase-9/3 indicated that the Fr-2 fraction induces apoptosis via mitochondria-dependent pathway in liver L-02 cells.

关 键 词：紫菀 肽类组分 肝细胞毒性 氧化应激 细胞凋亡 

分 类 号：R965[医药卫生—药理学]