青藤碱诱导人红白血病细胞株K562凋亡的实验研究  

Experimental Study on Apoptosis of Human Erythroleukemia Cell Line K562 Induced by Sinomenine

作　　者：牟凤林[1] 陈华[1] 

机构地区：[1]重庆三峡医药高等专科学校,重庆404100

出　　处：《中国中医基础医学杂志》2017年第10期1387-1390,1396,共5页JOURNAL OF BASIC CHINESE MEDICINE

基　　金：重庆市基础与前沿研究计划项目(cstc2014jcyj A10049)-构建I型大麻受体真核表达体系制备细胞膜色谱筛选中药中配体的研究;重庆市社会民生科技创新专项细胞色谱筛选新药技术用于中草药研发的示范应用(cstc2016shmszx130028);第二批重庆市高等学校青年骨干教师资助计划项目(2014)

摘　　要：目的:通过青藤碱(Sinomeine,SIN)诱导K562细胞凋亡实验,研究SIN诱导K562细胞凋亡机制。方法:CCK-8检测细胞增殖,流式细胞术(FMC)检测细胞凋亡,Hoechst 33258染色细胞凋亡形态学改变,Real-time PCR检测bcl-2和bax基因表达,Western blot检测Bcl-2、Bax、Caspase-3蛋白表达。结果:CCK-8和FCM结果显示,SIN可抑制K562细胞增殖并诱导其早期凋亡;Hoechst 33258染色结果显示,经SIN诱导48h后表现出细胞凋亡的典型变化;Real-time PCR结果显示,SIN组bax基因表达升高,bcl-2基因表达降低;Western blot结果显示,SIN组中Caspase-3、Bax蛋白表达升高,Bcl-2蛋白表达降低。结论:SIN可诱导K562病细胞凋亡从而抑制细胞增殖。Objective： To grope preliminarily the effects of Sinomenine（SIN） on the proliferation inhibition, apoptosis of human erythroleukemia cell line K562. Methods： CCK-8 assay was used to detect the K562 cell proliferation activity. K562 cell apoptotic changes were detected by Flow cytometry （FCM）. The cell morphological changes of apoptosis were observed by hoechst staining. The changes of expressions of bcl-2 gene and bax gene were examined by Real-time PCR. The changes of expressions of Bcl-2,Bax and Caspase-3 proteins were examined by western blot analysis. Results： CCK-8 assay result showed that SIN can effectively inhibit the proliferation of K562 cells in vitro, which exhibits a dose-dependent manner. Flow cytometry results indicated that SIN can induce early apoptosis of K562 cells after induced by SIN, which showed statistically significant difference compared with the control group. Hoechst staining revealed that SIN could induce toypical apoptotie morphological changes. Real-time PCR showed that the expressions of bax gene was increased in dose- dependent manner after induced by SIN, while expressions of bcl-2 gene were decreased compared with the control group. Western blot showed that the expressions of Bax, Caspase-3 were increased in dose-dependent manner after induced by SIN, expressions of Bcl-2 proteins were decreased compared with the control group. Conclusion： SIN can inhibit the proliferation of K562 cells and induce its apoptosis through activating Caspase-3 up-regulating the expression of Bax, and down-regulating the expression of Bax.

关 键 词：青藤碱 K562细胞 BCL-2 BAX 细胞凋亡 

分 类 号：R285.5[医药卫生—中药学]