齐墩果酸诱导Jurkat细胞凋亡及对ROS和细胞内Ca2+含量的影响

目的 探讨齐墩果酸对白血病Jurkat细胞株增殖抑制和诱导凋亡作用,分析凋亡发生与细胞内活性氧(ROS)及钙离子浓度(Ca2+]i)变化的关系.方法 应用MTT法检测细胞增殖抑制；流式细胞仪检测细胞凋亡率和细胞周期；DCFH-DA荧光定量法检测ROS含量；Fluo-3AM荧光负载方法测定Ca2+]i的变化.结果 齐墩果酸对Jurkat细胞的增殖具有浓度依赖性和时间依赖性抑制作用,0,40,80,160 μmol/L齐墩果酸作用24 h细胞凋亡率分别为(7.36±0.40)％、(19.80±1.59)％、(29.39±0.64)％、(34.72±0.94)％,G0/G1期细胞比例分别为(54.26±1.43)％、(85.83±0.91)％、(91.18±1.32)％、(92.90±1.19)％.80 μmol/L和160μmol/L齐墩果酸处理24 h,细胞中ROS和Ca2+]i水平均明显高于对照组(P＜0.05),ROS和Ca2+]i水平均与细胞凋亡率呈正相关(r分别为0.95、0.97).结论 细胞中ROS和Ca2+可能参与了齐墩果酸对Jurkat细胞的抑制增殖和诱导凋亡作用.

Oleanolic Acid Introduced Apoptosis in Jurkat Cells and Influence on the Contents of ROS and Intracellular Ca2+

Objective To investigate the effect of oleanolic acid(OA) on proliferative inhibition and apoptotic induction in Jurkat cells,and explore the relationship between OA-induced apoptosis and the change of intracellular ROS and Ca2+ concentration.Methods The cell inhibitory rate was assessed with MTT assay.The apoptotic rate and the cycle phase of Jurkat cells were measured with flow cytometry.ROS level was observed by DCFH-DA fluorescent quantitation,while intracellular Ca2+ concentration was measured using Fluo-3AM load method.Results The proliferation of Jurkat cells was inhibited by OA in a dose-and time-dependent manner.The apoptotic rates of Jurkat cells treated with OA(0,40,80 and 160 μmol/L) for 24 h were(7.36±0.40)%,(19.80±1.59)%,(29.39±0.64)% and(34.72±0.94)% and the cells of G0/G1 phase were(54.26±1.43)%,(85.83±0.91)%,(91.18±1.32)% and(92.90±1.19)%.ROS and intracellular Ca2+ concentration of treatment with 80 μmol/L and 160 μmol/L OA were higher than control(P < 0.05) and were positive correlated with apoptotic rates of Jurkat cells(r =0.95 and r =0.97).Conclusion The increase of ROS and intracellular Ca2+ might be involved in oleanlic acid-induced Jurkat cells apoptosis.