| 线粒体途径在大黄素促人混合培养淋巴细胞凋亡中的作用 |
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| 引用本文： | 沈乃营1,刘 昌2,何盟国1,曲 凯2,张天政1. 线粒体途径在大黄素促人混合培养淋巴细胞凋亡中的作用[J]. 现代肿瘤医学, 2015, 0(11): 1495-1498. DOI: 10.3969/j.issn.1672-4992.2015.11.06 |
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| 作者姓名： | 沈乃营1 刘 昌2 何盟国1 曲 凯2 张天政1 |
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| 作者单位： | 1.陕西核工业215医院肝胆外科，陕西 咸阳 712000；2.西安交通大学第一附属医院肝胆外科，陕西 西安 710061 |
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| 摘 要： | 目的:探讨大黄素促进人混合培养淋巴细胞凋亡的作用机制，为进一步研究大黄素的免疫抑制作用奠定实验基础。方法:建立混合淋巴细胞培养模型，给予不同浓度大黄素（1μmol/L、10μmol/L、100μmol/L）培养24、48、72h后检测各组淋巴细胞凋亡、细胞内活性氧及线粒体膜电位情况。Western blot法检测共培养48h后淋巴细胞中细胞色素C、caspase-3、caspase-9、Bcl-2/Bax蛋白表达变化。给予还原性谷胱甘肽（GSH）清除细胞内活性氧后，检测其对线粒体膜电位及淋巴细胞凋亡的影响。结果:随着培养时间和药物浓度的增加，大黄素对淋巴细胞增殖抑制和促凋亡作用逐渐增强，呈现浓度和时间依赖性。高浓度大黄素提高细胞内活性氧、降低线粒体膜电位水平，激活线粒体凋亡通路。使用还原性谷胱甘肽清除细胞内活性氧后，能够降低线粒体膜电位和淋巴细胞凋亡率。结论:线粒体信号途径的激活在大黄素促淋巴细胞凋亡中发挥重要作用。
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| 关 键 词： | 大黄素 ROS 细胞色素C caspase蛋白 凋亡 Bcl-2家族 淋巴细胞 |
Effect of the mitochondrial signaling pathway on Emodin-induced apoptosis in human mixed lymphocyte culture |
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| Shen Naiying1,Liu Chang2,He Mengguo1,Qu Kai2,Zhang Tianzheng1. Effect of the mitochondrial signaling pathway on Emodin-induced apoptosis in human mixed lymphocyte culture[J]. Journal of Modern Oncology, 2015, 0(11): 1495-1498. DOI: 10.3969/j.issn.1672-4992.2015.11.06 |
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| Authors: | Shen Naiying1 Liu Chang2 He Mengguo1 Qu Kai2 Zhang Tianzheng1 |
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| Affiliation: | 1.Department of Hepatobiliary Surgery，Shaanxi Xianyang 215 Hospital,Shaanxi Xianyang 712000，China;2.Department of Hepatobiliary Surgery，the First Affiliated Hospital Medical School of Xi'an Jiaotong University，Shaanxi Xi'an 710061,China. |
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| Abstract: | Objective:To investigate the effect of mitochondrial signaling pathway on Emodin-induced apoptosis in human mixed lymphocyte culture model.Methods:The model of human mixed lymphocyte culture was established and lymphocytes were isolated from the peripheral blood samples of different healthy adult volunteers.To investigate the immunosuppressive effect of Emodin.Emodin was administered with different concentrations at 1μmol/L,10μmol/L,100μmol/L for 24,48,72h.At the end of culture，the apoptotic rate of lymphocytes and the level of reactive oxygen species （ROS） and mitochondrial membrane potential （ΔΨm） were evaluated with flow cytometry.Protein expressions of cytosolic cytochrome C，caspase-3 and caspase-9 were analyzed by Western blot.In the high concentration Emodin group，lymphocytes were cultured with reduced glutathione （GSH） and Emodin for 48h，GSH can remove intracellular ROS.The apoptotic rate of lymphocytes and the levels of ROS and ΔΨm were also evaluated by flow cytometry.Protein expressions of Bcl-2 and Bax were analyzed by Western blot.Results:With the enhanced concentration of Emodin，the inhibition of proliferation and increase of apoptotic rate were observed.The results further showed generation of ROS，disruption of ΔΨm，decrease of the Bcl-2/Bax ratio and cytosolic cytochrome C.Caspase-3 and caspase-9 levels also increased after the Emodin treatment.The decline in the Bcl-2/Bax ratio and the increase of ROS,cytosolic cytochrome C and caspase-3 and caspase-9 levels were consistent with the increase of the lymphocyte apoptotic ratio.In the GSH group，the level of ROS declined，and the apoptotic rate also reduced compared with the high concentration Emodin group （P<0.05)，however，the ΔΨm enhanced compared with control group （P<0.05).Conclusion:Our results strongly suggested that the mitochondrial signaling pathway was involved in Emodin-induced apoptosis in human mixed lymphocyte culture. |
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| Keywords: | Emodin ROS cytochrome C caspase protein apoptosis Bcl-2 lymphocyte |
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