摘要
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,0,40,80,160μmol/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)%。80μmol/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。
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.
出处
《中国医科大学学报》
CAS
CSCD
北大核心
2012年第10期921-925,共5页
Journal of China Medical University
基金
辽宁省科学技术计划(2010225032)
