黄芩苷通过诱导ROS发生抑制人T细胞淋巴瘤的增殖并促进凋亡

Baicalin inhibits the proliferation and promotes apoptosis of human T-lymphoma Molt4 cells by inducing ROS production

目的:研究黄芩苷(baicalin)对T细胞淋巴瘤Molt4细胞增殖的抑制作用,并探讨其可能的作用机制。方法:采用不同质量浓度的黄芩苷(2.5、5、10、20、40、80和160μg/mL)处理T细胞淋巴瘤Molt4细胞,MTT法检测细胞增殖抑制率并计算半数抑制浓度(half maximal inhibitory concentration,IC50)值。用黄芩苷(20μg/mL)处理Molt4细胞后,采用瑞氏染色法检测对细胞凋亡形态的影响,FCM法检测对Molt4细胞凋亡率的影响,DCFH-DA法检测Molt4细胞中活性氧(reactive oxygen species,ROS)含量的变化。最后,采用FCM法检测黄芩苷单药或联合二硫苏糖醇(dithiothreitol,DTT)对Molt4细胞凋亡的影响。结果:不同质量浓度的黄芩苷(2.5、5、10、20、40、80和160μg/mL)可明显抑制Molt4细胞的增殖能力(P值均<0.05),黄芩苷对Molt4细胞的IC50值为(19.2±2.2)μg/mL。黄芩苷处理Molt4细胞48和60 h后均出现明显的凋亡形态;FCM法检测显示黄芩苷组细胞的凋亡率为(42.7±5.0)%,明显高于对照组的(5.0±0.9)%(P<0.01);DCFH-DA法检测结果显示黄芩苷处理Molt4细胞6、12和24 h后细胞内ROS的水平随时间的延长而明显提高,但在36 h后细胞内ROS的水平开始降低。黄芩苷联合DTT可明显降低黄芩苷对Molt4细胞凋亡率(P<0.01)。结论:黄芩苷可抑制T淋巴瘤Molt4细胞的生长,并通过刺激ROS生成从而诱导Molt4细胞的凋亡。

Objective:To investigate the effect of baicalin on the growth of T-lymphoma Molt4 cells,and it’s possible mechanism.Methods:T-lymphoma Molt4 cells were treated with different concentrations baicalin(2.5,5,10,20,40,80 and 160μg/mL),the cell growth inhibitory rate was detected by MTT method and the half maximal inhibitory concentration(IC50)was calculated.T-lymphoma Molt4 cells were treated with baicalin(20μg/mL),the morphological characteristic of apoptosis was observed by light microscopy after Wright’s staining.The level of reactive oxygen species(ROS)in Molt4 cells was detected by 2’,7’-dichlorodihydrofluorescein diacetate(DCFH-DA)staining.The apoptosis rate of Molt4 cells treated with baicalin alone or baicalin combined with dithiothreitol(DTT)was detected by FCM method.Results:After treatment with different concentrations baicalin(2.5,5,10,20,40,80 and 160μg/mL)for 72 h,baicalin could inhibit the cell proliferation of Molt4 cells(all P<0.05),the IC50 value of baicalin was(19.2±2.2)μg/mL.The obvious apoptosis morphology of Molt4 cells was observed after treatment with baicalin.The cell apoptosis rate in the 20μg/mL baicalin group[(42.7±5.0)%]was significantly higher than the control group[(5.0±0.9)%](P<0.01).The ROS level was increased after 6,12 and 24 h in the baicalin group,the ROS level was time-dependent,and the ROS level was decreased after 36 h.Baicalin combined with DTT could significantly reduce the apoptosis rate of Molt4 cells induced by baicalin(P<0.01).Conclusion:Baicalin can inhibite the growth of T-lymphoma Molt4 cells,proliferation by cause apoptosis via stimulating ROS production.