白花蛇舌草对人胶质瘤细胞增殖及凋亡的影响及其机制

Effects of hedyotic diffusa on human glioma cells proliferation and apoptosis and the mechanism

目的 观察白花蛇舌草在体外对胶质瘤U87细胞株增殖和凋亡的影响并探讨其机制.方法 用不同浓度的白花蛇舌草含药培养基（0、5、10、15、20ml/L）处理U87细胞不同时间（24、72 h）后,采用噻唑蓝（MTT）比色法检测其对U87细胞增殖能力的影响,流式细胞仪检测对照组与实验组细胞的凋亡差异,免疫印迹法检测U87细胞中半胱氨酸蛋白酶（Caspase-3）表达水平的改变.结果 经白花蛇舌草5、10、15、20ml/L处理后,U87细胞增殖受到明显抑制,呈显著的剂量依赖性 经白花蛇舌草（5、10 ml/L）处理后U87细胞的凋亡率分别增加到（5.22±0.29）%、（8.38±0.65）%,明显高于对照组的（3.09±0.05）%（P＜0.01） 白花蛇舌草可明显上调U87细胞中Caspase-3的蛋白表达,且呈浓度依赖性.讨论白花蛇舌草对胶质瘤U87细胞生长的抑制作用可能通过上调Caspase-3基因的表达从而诱导细胞凋亡实现.

Objective To investigate the effects of hedyotic diffusa （HD） on glioma U87 cells proliferation and apoptosis and the possible mechanism. Methods U87 cells were cultured for 24 and 72 h in medium containing HI） with different concentrations （0, 5, 10, 15, 20 ml/L）. The methylthiazol tetrazolium （MTT） assay was used to detect the survival rate, the apoptosis of the cells was examined by flow cytometry, and the change of caspase-3 protein level was measured by Western blotting. Results The cells proliferation was obviously inhibited by HD in a dose-dependent manner. The apoptosis rate in 5, 10 ml/L HD groups was （5.22 ± 0. 29） % and （ 8. 38 ± 0.65 ） % respectively, which was significantly higher than that in 0 ml/L HD group （P 〈 0. 01 ）. The protein expression of caspase-3 was increased dose-dependently. Conclusion The growth inhibition of glioma U87 cells caused by HD may be associated with the upregulation of caspase-3 gene, which induced apoptosis.