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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 of polysaccharide Saponin Ⅱ on the apoptosis of melanoma B16 cells and its mechanism. METHODS: MTT assay was used to observe the inhibitory effect of polysaccharide Saponin Ⅱ on the proliferation of B16 cells. The morphological changes of B16 cells induced by polysaccharide Saponin Ⅱ were observed under inverted fluorescence microscope with DAPI staining. Annexin V-FITC / PI double staining And flow cytometry were used to detect the effect of polysaccharide Saponin Ⅱ on the apoptosis rate of B16 cells; JC-1 staining flow cytometry was used to detect the changes of mitochondrial membrane potential of B16 cells treated with Polypeptide Saponins Ⅱ; On the expression of Bcl-2 in B16 cells. Results: Pollen saponin Ⅱ inhibited the proliferation of B16 cells in a time and concentration-dependent manner. The apoptotic rates of B16 cells treated with polysaccharide saponin Ⅱ increased with the increase of drug concentration. The mitochondrial membrane potential (P <0.01). The results of Western blot showed that the expression of Bcl-2 protein was significantly lower than that of the blank control group (P <0.01). Conclusion: Tail saponin Ⅱ may induce B16 cell apoptosis through mitochondrial oxidative stress pathway.