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

Changes of mitochondrial membrane potential and mitochondrial mass in camptothecin-induced Jurkat cells

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-1CPT诱导下,6 h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G0/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,AnnexinV+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。

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.