线粒体通路和死亡受体通路在中华眼镜蛇毒组分诱导KG1a细胞凋亡中的作用  

The action of mitochondrial apoptosis pathway and death receptor pathway in the process of Naja Naja Actra Venom Component inducing the KG1a apoptosis

作　　者：贺艳杰[1] 李玉华[1] 卢会芳[1] 黄宇贤[1] 黄睿[1] 郭坤元[1] 

机构地区：[1]南方医科大学珠江医院血液科,广东广州510282

出　　处：《中国药理学通报》2013年第3期356-360,共5页Chinese Pharmacological Bulletin

基　　金：国家自然科学基金资助项目(No 30973454)

摘　　要：目的研究线粒体通路和死亡受体通路在中华眼镜蛇毒组分(naja naja actra venom component,NNAVC)诱导KG1a细胞凋亡过程中的作用。方法以人急性髓系白血病细胞株KG1a细胞为研究对象,采用Annexin V-FITC/PI荧光染色法进行凋亡细胞的形态学观察,流式细胞仪检测细胞凋亡率,分光光度法检测Caspase-9、Caspase-8和Caspase-3的活性,ELISA法检测胞质内细胞色素C的表达水平,流式细胞仪分析TRAIL死亡受体(DR4、DR5)和诱骗受体(DcR1、DcR2)的改变。结果 NNAVC作用后,荧光显微镜下可见明显的凋亡细胞,细胞凋亡率随药物作用浓度的增加而升高;Caspase-9、Caspase-8和Caspase-3蛋白被激活,胞质内细胞色素C的表达明显增加;流式细胞仪检测结果显示,NNAVC具有降低死亡受体DR4和诱骗受体DcR1的表达率,而提高DR5和DcR2的表达水平的作用。结论线粒体凋亡通路和死亡受体通路均参与NNAVC诱导KG1a细胞凋亡的过程,这可能是NNAVC发挥抗白血病作用的机制之一。Aim To investigate the action of death receptor pathway and mitochondrial apoptosis pathway in the process of NNAVC（Naja Naja Actra Venom Component） inducing the KG1a apoptosis.Methods The human acute myeloid leukemia（AML） cell KG1a was chosen as experimental subject,Annexin V-FITC/PI fluorescent staining was used to observe the morphological changes of apoptotic cells,and the apoptosis rates were measured by flow cytometry;the activities of caspase-9,caspase-8 and caspase-3 were measured by colorimetric method;cytochrome C level was measured by ELISA assay;TRAIL DR4,DR5 and DcR1,DcR2 were analyzed by flow cytometry.Results The apoptotic cells could be seen clearly under fluorescence microscope,the apoptotic rates of KG1a cells were raised along with NNAVC′s concentration;caspase-9,caspase-8 and caspase-3 were elevated after the treatment;cytochrome C level increased noticeably compared with than in control group;the analysis showed that the protein expression of DR4 and DcR1 on KG1a cells was decreased,but the expression rate of DR5 and DcR2 was increased according to flow cytometry.Conclusion Both mitochondrial apoptosis pathway and death receptor pathway are involved in the apoptosis process of KG1a cells induced by NNAVC,which may be one of the antitumor mechanisms of NNAVC.

关 键 词：白血病 中华眼镜蛇毒 KG1a细胞 线粒体通路 死亡受体通路 凋亡 

分 类 号：R329.25[医药卫生—人体解剖和组织胚胎学] R392.11[医药卫生—基础医学]