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目的:研究莪术醇对小鼠皮肤黑色素瘤B16-F10细胞凋亡的影响。方法:采用四氮唑盐(MTT)法检测不同浓度的莪术醇对B16-F10细胞增殖的影响;并用光学显微镜观察不同浓度的莪术醇对B16-F10细胞状态的影响;通过流式FITC-Annexin V/PI双染法,检测不同浓度的莪术醇对细胞凋亡率的影响;通过间接荧光标记法检测不同浓度的莪术醇对细胞内活性氧水平的影响;通过罗丹明123染色,检测不同浓度的莪术醇对细胞内线粒体膜电位的影响。结果:不同浓度莪术醇处理B16-F10细胞24、48、72h,均可呈浓度、时间依赖性的抑制细胞增殖;显微镜观察到莪术醇可以引起细胞形态变圆,漂浮等细胞死亡现象。12.5、25、50、100μg/ml莪术醇作用48h后,诱导的细胞凋亡率分别为(19.5±3.4)%、(35.1±2.8)%、(45.9±4.1)%、(60.5±1.9)%,与对照组具有显著性统计学差异;诱导的细胞内活性氧水平的百分率分别为(6.9±1.8)%、(12.4±2.1)%、(20.9±3.1)%、(29.1±3.5)%,与对照组相比具有显著统计学差异。引起的线粒体膜电位的百分比分别为(40±1.1)%、(33.7±4.2)%、(27.3±2.5)%、(17.9±2.9)%,与空白对照组相比具有统计学差异。结论:莪术醇可以抑制B16-F10的增殖并促进细胞凋亡,其凋亡机制可能与细胞内活性氧组分的产生以及线粒体膜电位的变化有关。
Objective: To study the effect of curcumol on the apoptosis of mouse skin melanoma B16-F10 cells. Methods: The effects of curcumol at different concentrations on the proliferation of B16-F10 cells were detected by MTT assay. The effects of curcumol at different concentrations on the cell cycle of B16-F10 cells were observed by light microscopy. V / PI double staining method was used to detect the effect of different concentrations of curcumol on the apoptosis rate; the indirect fluorescent labeling method was used to detect the effects of different concentrations of curcumol on intracellular reactive oxygen species; rhodamine 123 staining, Effect of curcumol on mitochondrial membrane potential in cells. Results: Curcumol treatment of B16-F10 cells for 24, 48, and 72 h could inhibit cell proliferation in a concentration-dependent and time-dependent manner. Curcumol could cause cell morphology such as round and floating cells under microscope. The apoptosis rates induced by 12.5, 25, 50, 50μg / ml Curcumol were (19.5 ± 3.4)%, (35.1 ± 2.8)%, (45.9 ± 4.1)%, (60.5 ± 1.9)% (6.9 ± 1.8)%, (12.4 ± 2.1)%, (20.9 ± 3.1)% and (29.1 ± 3.5)%, respectively, which were significantly different from the control group There was a significant statistical difference compared with the control group. (40 ± 1.1)%, (33.7 ± 4.2)%, (27.3 ± 2.5)% and (17.9 ± 2.9)%, respectively, which were significantly different from those of the blank control group. CONCLUSION: Curcumol can inhibit the proliferation of B16-F10 cells and promote apoptosis. The mechanism of apoptosis may be related to the generation of intracellular reactive oxygen species and changes of mitochondrial membrane potential.