【摘要】 目的探讨香叶木素对人乳腺癌MCF-7细胞增殖、凋亡的影响及其作用机制。方法将MCF-7细胞分为对照组(0μmol·L^-1)和香叶木素5、10、20、30、40、50、60、70μmol·L^-1浓度组。采用CCK-8法及2,4二硝基苯肼法检测MCF-7细胞活力、乳酸脱氢酶（LDH）泄露量;采用罗丹明123（Rh123）荧光染色法、流式细胞术及荧光显微镜检测MCF-7细胞线粒体膜电位（MMP）;DCFH-DA法检测细胞内活性氧（ROS）含量;Annexin V-FITC/PI双染法检测细胞凋亡;Western Blot法检测p53、Bax、Caspase 3及Bcl-2蛋白的表达。结果与对照组比较,香叶木素在5～70μmol·L^-1范围内能够浓度依赖性地抑制MCF-7的细胞活力及促进LDH的泄露（P<0.01,P<0.001）,选取浓度10、30、50μmol·L^-1进行后续实验;香叶木素10、30、50μmol·L^-1浓度组能显著降低MCF-7细胞线粒体膜电位（P<0.05,P<0.01,P<0.001）,促进细胞内ROS的积累（P<0.01,P<0.001）,提高细胞凋亡率（P<0.001）,显著上调p53(30、50μmol·L^-1)、Bax及Caspase 3蛋白的表达（P<0.001）,下调Bcl-2蛋白的表达（P<0.01,P<0.001）。结论香叶木素可能通过ROS及p53介导的线粒体凋亡途径抑制MCF-7细胞的增殖及促进其凋亡。更多还原

【Abstract】 Objective To study the anti-proliferation and pro-apoptotic effects of diosmetin on MCF-7 cells and elucidate the underlying molecular mechanisms.Methods The MCF-7 cells were divided into control group(0 μmol·L^-1)and diosmetin 5,10,20,30,40,50,60 and 70 μmol·L^-1 groups.The effects of diosmetin on cell viability and LDH leakage of MCF-7 cells were assessed using CCK-8 and an LDH assay kit.The effect of diosmetin on mitochondrial membrane potential（MMP） of MCF-7 cells was measured by Rhodamine 123（Rh123） fluorescence staining and flow cytometry and a fluorescence microscope.The effects of diosmetin on the level of reactive oxygen species（ROS） of MCF-7 cells was evaluated using DCFH-DA fluorescence staining and flow cytometry.The effects of diosmetin on cell apoptosis of MCF-7 cells was analysed using an annexin V-FITC/PI assay kit and flow cytometry.Western Blot was used to detect the expression levels of p53,Bax,Bcl-2 and Caspase 3 proteins in MCF-7 cells.Results Compared with the control group,the cell viabilities of MCF-7 cells treated with diosmetin were decreased and LDH release was increased in a dose-dependent manner（P<0.01,P <0.001）.And the groups treated with 10,30,50 μmol·L^-1 diosmetin could significantly decrease the MMP of MCF-7 cells（P <0.05,P <0.01,P<0.001）,increase the intracellular ROS（P <0.01,P <0.001） and promote the apoptosis of MCF-7 cells（P <0.001）.The protein expression levels of p53,Bax,and Caspase 3 were up-regulated significantly in diosmetin10,30,50 μmol·L^-1 groups（P <0.001）;while the expression levels of Bcl-2 protein were down-regulated significantly（P <0.01,P <0.001）.Conclusion Diosmetin exhibited anti-proliferative and pro-apoptotic effects on MCF-7 cells through mitochondrial apoptosis pathway mediated by ROS and p53.更多还原