重楼皂苷Ⅱ诱导黑色素瘤B16细胞凋亡的机制研究  

Study on the Mechanism of Polyphyllin Ⅱ Induced Apoptosis of Melanoma B16 Cells

作　　者：程卉[1] 苏婧婧[1] 王训翠[1] 李庆林[1] CHENG Hui SU Jing-jing WANG Xun-eui LI Qing-lin(Anhui University of Traditional Medicine, Key Laboratory of Xin＇an Medicine, Ministry of Education, Hetei 230038, Chin)

机构地区：[1]安徽中医药大学/教育部新安医学重点实验室,安徽合肥230038

出　　处：《中药材》2016年第11期2594-2597,共4页Journal of Chinese Medicinal Materials

基　　金：国家自然科学基金面上项目(81173600;81673650)

摘　　要：目的:研究重楼皂苷Ⅱ诱导黑色素瘤B16细胞凋亡的作用及机制。方法:采用MTT法观察重楼皂苷Ⅱ对B16细胞增殖的抑制作用;采用DAPI染色于倒置荧光显微镜下观察重楼皂苷Ⅱ对B16细胞诱导凋亡的形态学改变;Annexin V-FITC/PI双染及流式细胞术检测重楼皂苷Ⅱ对B16细胞凋亡率的影响;JC-1染色流式细胞仪检测重楼皂苷Ⅱ作用B16细胞后细胞线粒体膜电位的变化;Western blot检测重楼皂苷Ⅱ对B16细胞中凋亡相关蛋白Bcl-2的表达变化。结果:重楼皂苷Ⅱ对B16细胞的增殖有明显的抑制作用,并呈时间及浓度依赖性;重楼皂苷Ⅱ作用B16细胞后凋亡率随药物浓度的升高而升高;线粒体膜电位水平也随之显著降低,与空白对照组比较差异有统计学意义(P<0.01);Western blot检测结果表明Bcl-2蛋白表达水平显著降低。结论:重楼皂苷Ⅱ可能通过线粒体氧化应激通路从而诱导B16细胞凋亡。Objective ：To study the effect and mechanism of Polyphyllin R inducing B16 cell apoptosis, Methods：MTT assay was used to detect B16 cell vitality rate after treated with PolyphyllinⅡ;DAPI staining was used to observe the B16 cell morphological change after treated with Polyphyllin Ⅱ by Fluorescence microscope;Annexin V-FITC/PI staining and Flow cytometry analysis were used to determine the apoptosis rate of Polyphyllin Ⅱ which influenced on B16 cell;JC-1 was used to detect the B16 cell mitochondrial membrane potential changes after treated with Polyphyllin Ⅱ;the protein expression change of Bcl-2 was detected hy Western blotting. Resuhs：Polyphyllin Ⅱ can inhibit B16 cell proliferation in a time and concentration dependant manner significantly; compared with control group,the B16 cell apoptosis rate increased as the concentration of Polyphyllin 11 increased while mitochondrial membrane potential decreased;Western blotting test results indicated that the expression of Bcl-2 decreased significantly. Conclusion： Polyphyllin Ⅱ can induce B16 cell apoptosis through Mitochondrial oxidative stress pathways.

关 键 词：重楼皂苷Ⅱ 黑色素瘤B16细胞 线粒体氧化应激 细胞凋亡 

分 类 号：R285.5[医药卫生—中药学]