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-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.
出处
《中国病理生理杂志》
CAS
CSCD
北大核心
2006年第5期846-850,共5页
Chinese Journal of Pathophysiology
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)