白花蛇舌草对人B淋巴细胞瘤Ramos细胞增殖和凋亡的影响

Effect of Herba Hedyotis Diffusae on Ramos cell proliferation and apoptosis

目的探讨白花蛇舌草对Ramos细胞增殖和凋亡的影响。方法使用0、10、20、40 m L·L-1白花蛇舌草处理Ramos细胞24 h后收获细胞;采用细胞增殖检测试剂盒检测白花蛇舌草对Ramos细胞增殖的影响,并检测Ramos细胞线粒体膜电位的变化;流式细胞术检测白花蛇舌草对Ramos细胞凋亡的影响,并检测Ramos细胞中caspase-3、caspase-9活性的变化;Western blot法检测Ramos细胞Bax、Bcl-2蛋白的表达。结果细胞增殖实验结果证明白花蛇舌草处理的Ramos细胞增殖受到明显抑制;20、40 m L·L-1白花蛇舌草干预Ramos细胞24 h后,Ramos细胞线粒体膜电位低于0 m L·L-1(P<0.05,P<0.01);流式细胞术检测结果表明细胞凋亡增加;与0 m L·L-1白花蛇舌草比较,20 m L·L-1白花蛇舌草处理Ramos细胞24 h,caspase-3和caspase-9活性增加了96%和37%(P<0.05),40 m L·L-1白花蛇舌草处理Ramos细胞24 h,caspase-3和caspase-9活性增加了148%和115%(P<0.01)。Western blot法证实Bax蛋白表达量增强,而Bcl-2蛋白表达量减弱,且呈现一定程度的剂量依赖性。结论白花蛇舌草可通过活化调控胱天蛋白依赖的线粒体途径,诱导Ramos细胞凋亡。

Objective To investigate the effect of Herba Hedyotis Diffusae（ HHD） on proliferation and apoptosis of Ramos cells. Methods Ramos cell was intervened by 0,10,20,40 m L·L- 1HHD,then the cell was harvested after 24 h.Cell proliferation was measured with Cell Counting Kit 8（ CCK8） assay;and the membrane potentials of mitochondria was detected. Cell apoptosis was detected with flow cytometry;the activity of caspase-3 and caspase-9 was detected. The expression of Bax and Bcl-2 was analyzed by Western blot. Results CCK8 assay results showed that HHD had inhibitory effect on Ramos cells. The membrane potentials of mitochondria of Ramos cells which was intervened by 20,40 m L·L- 1HHD for 24 h,was lower than that was intervened by 0 m L·L- 1（P 0. 05,P 0. 01）. Flow cytometry revealed that apoptotic rate of Ramos cells increased. Compared with 0 m L·L- 1HHD,the activity of caspase-3 and caspase-9 in Ramos cells which was intervened by 20 m L·L- 1HHD for 24 h increased by 96% and 37%（ P 0. 05）,while it increased by 148% and 115% which was intervened by 40 m L·L- 1HHD for 24 h（ P 0. 01）. Furthermore,the result of Western blot indicated that the Bax protein expression increased while Bcl-2 protein expression decreased in a dose-dependent manner. Conclusion HHD can induce apoptosis of Ramos,the molecular mechanism underlying this might involve the modulation of mitochondrial pathway.