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喜树碱诱导Jurkat细胞线粒体膜电势和线粒体质量变化的研究

ISSN号：1000-4718
期刊名称：《中国病理生理杂志》
时间：0
分类：R979.1[医药卫生—药品;医药卫生—药学] R363[医药卫生—病理学;医药卫生—基础医学]
作者机构：[1]暨南大学教育部组织移植与免疫重点实验室,广东广州510632
相关基金：国家自然科学基金重点项目（No.30230350）;国家自然科学基金面上项目（No.30471635）;广东省自然科学基金资助项目（No.5300413）;暨南大学引进优秀人才科研启动基金资助项目（No.51204066）

作者：江颖娟[1], 曾耀英[1], 王通[1], 肇静娴[1], 刑飞跃[1], 王希朝[1], 梁佩燕[1]

关键词：喜树碱, JURKAT细胞, 线粒体, 膜电位, 细胞凋亡, Camptothecin, Jurkat cells, Mitochondrial, Membrane potentials, Apoptosis

中文摘要：

目的：研究喜树碱（camptothecin，CPT）诱导Jurkat细胞凋亡过程中线粒体膜电势和线粒体质量的变化。方法：用喜树碱处理Jurkat细胞，利用Annexin V-FITC／PI双染流式细胞术研究细胞早期凋亡，PI染色流式细胞术测细胞周期，Annexin V-PE／DiOC6（3）双染流式细胞术检测线粒体膜电势（△ψm），NAO染色流式细胞术检测线粒体质量。结果：在10μmol·L^-1 CPT诱导下，6h时Jurkat细胞早期凋亡的细胞比率（22．59±1．04）％显著高于对照组（3．93±0．73）％（P〈0．01）。CPT组坏死比率（2．48±0．53）％与对照组（2．78±0．63）％无显著差异（P〉0．05）；并可使细胞出现明显的凋亡峰。晚期凋亡的细胞比率为（13．58±0．97）％显著高于对照组（3．18±0．51）％（P〈0．01），CPT组C0／G1期细胞比率（48．14±0．96）％，明显高于对照组（44．09±0．43）％（P〈0．01）。CPT组线粒体发生明显去极化现象，Annexin V^＋DiOC6（3）-的细胞比率为（19．47±0．69）％，而对照组比率为（4．21±0．40）％，差异显著（P〈0．01）。同时，CPT组线粒体质量显著低于对照组：CPT组NAO^＋细胞比率为（74．77±1．66）％，对照组为（92．24±1．41）％（P〈0．01）。结论：CPT诱导Jurkat细胞凋亡过程中线粒体去极化作用增强并且线粒体质量下降，表明该凋亡过程与线粒体途径密切相关。

英文摘要：

AIM： To study the changes of mitochondrial membrane potential （△ψm） and mitochondrial mass in apoptosis of Jurkat cells induced by camptothecin （CPT）. METHODS： Jurkat cells were treated with CPT. Annexin V - FITC/propidium iodine （PI） double stainig was used,to detected early stage of apoptosis and PI staining for analyzing the cell cycle. Jurkat cells were stained by annexin V - PE/DiOC6（3） to detect changes of △ψm. The mitochondrial mass was measured by cytometry with NAO staining. RESULTS： 6 h afar treated with 10 μmol/L CPT, the rate of early apoptotic cells （22.59± 1.04）% had significantly difference compared with control group （3.93 ± 0. 73） % （ P 〈 0.01 ）. The necrotic rate （2.48 ±0. 53） % had no significant difference to that in control group （2.78 ± 0.63）% （ P 〉 0.05）. Apoptotic peak appeared obviously after treated with CPT, the percentage of late apoptotic cells （ 13.58 ± 0. 97） % had distinctly difference compared with control group （3.18 ± 0.51 ） % （ P 〈 0.01 ）. The cells in G0/G1 phase （48.14±0.96）% were much higher than that in control group （44.09±0.43）% （P〈0.01）. Mitochondrial depolarization was very obviously in CPT group. The percentage of annexin V^＋DiOC6（3） - cells was （ 19.47 ± 0.69） %, while in control group, was （4.21 ± 0.40） % （ P 〈 0.01 ）. Mitochondrial mass in CPT group was significantly lower than that in control group, the percentage of NAO^＋ cells （74.77 ± 1.66） % had significantly difference compared with control group （92.24 ± 1.41 ）% （P 〈 0.01 ）. CONCLUSION： During the process of CPT- induced apoPtosis in Jurkat cells, mitochondrial depolarization was very obviously and mitochondrial mass decreased, indicating that the process of apoptosis is nearly related to the mitochondrial pathway.

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