新藤黄酸诱导黑色素瘤B16细胞凋亡的研究  

Apoptosis of Melanoma B16 Cells Induced by Gambogenic Acid

作　　者：谈永进[1,2] 张璇[2] 程卉[2] 李庆林[1,2] 

机构地区：[1]安庆医药高等专科学校,安徽安庆246052 [2]安徽中医药大学科研实验中心,安徽合肥230038

出　　处：《中药材》2014年第3期469-473,共5页Journal of Chinese Medicinal Materials

基　　金：安徽省自然科学基金(11040606M190);国家自然科学基金(81173600);教育部优秀人才基金

摘　　要：目的:研究新藤黄酸(Gambogenic Acid,GNA)对黑色素瘤B16细胞的增殖抑制作用,并探讨细胞凋亡机制在其中的角色。方法:MTT法检测新藤黄酸对B16细胞生长和增殖的作用;采用acridine orange/ethidium bromide(AO/EB)荧光染色观察新藤黄酸对B16细胞的影响;采用流式细胞仪检测新藤黄酸处理B16细胞后细胞内活性氧(ROS)的产生;Western blotting法检测凋亡相关蛋白Caspase-3的变化。结果:MTT结果表明新藤黄酸在一定浓度下对黑色素瘤B16细胞的增殖有明显抑制作用,并呈一定的时效和量效关系;AO/EB染色观察表明新藤黄酸处理的细胞呈现明显的凋亡状态;新藤黄酸处理细胞后,细胞内活性氧的急剧增加,与未给药组比较有统计学意义(P<0.01),同时线粒体膜电位也随之下降;Western blotting检测发现新藤黄酸作用后cleaved-Caspase-3蛋白表达量增加。结论:新藤黄酸在一定的时间和浓度范围内能够抑制黑色素瘤B16细胞的增殖,诱导细胞发生凋亡。Objective：To study the inhibitory effect of Gambogenic acid（GNA） on melanoma B16 cells proliferation, and to explore the role of cell apoptosis. Methods ：The inhibitory effect of GNA on the proliferation of B16 cells was measured by methyl thiazolyl tetrazolium（MTY） assay;Alternation of B16 cells ultrastructure was detected by AO/EB staining under fluorescent microscope;Flow cytometry was used to detect intracellular reactive oxygen species （ROS）in B16 ceils generated by GNA treatment. Western blotting was used to investigate the expression of intracellular Caspase-3 proteins changes. Results ： MTF results showed that the GNA within a certain time and a certain concentration significantly suppressed the proliferation of B16 cells and morphological changes were observed by fluo- rescence microscope on B16 cells after GNA treatment. AO/EB staining showed that the major cell density decreased. GNA treated cells showed obvious apoptotic status. After the cells treated with GNA, in a short period of time,intracellular ROS levels increased dramatically compared with the control group（P 〈 0. 01 ）, and the mitochondrial membrane had a low potential consistently. Western blotting re- sults showed that changes of intracellular proteins expression in the release of Caspase-3 proteins expression levels were increased after GNA treatment. Conclusion：GNA can inhibit malignant melanoma B16 cells growth and proliferation and induce apoptosis within a certain time and at a certain concenlration.

关 键 词：新藤黄酸 细胞凋亡 黑色素瘤B16细胞 

分 类 号：R285.5[医药卫生—中药学]