高浓度谷氨酸对原代培养人脑胶质瘤细胞生长抑制和凋亡的影响

Influence of high concentration of glutamate on the inhibition of growth and apoptosis of primary cultured human glioma cells

目的探讨高浓度谷氨酸对人脑胶质瘤原代培养细胞的生长抑制和凋亡诱导作用。方法在体外对新鲜的人脑胶质瘤标本进行原代培养;不同浓度的谷氨酸作用不同时间后,进行形态学观察噻唑蓝(MTT)法检测生长抑制率;流式细胞仪Annexin V-FITC／PI双染法检测凋亡率。结果原代人脑胶质瘤细胞短期培养成功。高浓度谷氨酸对培养细胞有明显的抑制作用,并且存在一定的剂量依赖关系。当谷氨酸的浓度为25～50mmol／L时,可明显诱导培养细胞凋亡,并且凋亡率随药物浓度增加和时间延长而增高;当谷氨酸浓度达100 mmol／L以上时,细胞则主要以坏死为主。结论以酶消化法可成功对人脑胶质瘤细胞进行体外原代培养;在一定的浓度范围内,高浓度谷氨酸能明显的抑制原代培养人脑胶质瘤细胞生长,并诱导其凋亡。

Objective To study the induction of high concentration of glutamate on the growth inhibition and apoptosis of primary cultured human glioma cells in vitro. Methods Fresh tissues from dif- ferent gliomas were obtained to be cultured in vitro. After treatment with high concentration of glutamate, the morphologic changes were observed, the inhibitory effect was assessed by MTT and the apoptosis rate was quantified by flow cyctometry. Results The primary human glioma cells were successfully cultured. High concentration of glutamate inhibited the growth of cultured glioma cells in a dose-dependent manner. When the concentration of glutamate was 25-50 mmol/L, the apoptosis was induced and the apoptotic rate was increased with the concentration and time. But when the concentration was above 100 mmol/L,necrosis was the main pattern of cells death. Conclusion The primary human glioma cells were successfully cultured in vitro by enzymatic digestion culture method. High concentration of glutamate （2550 mmol/L） can obviously inhibit the growth of primary cultured glioma cells and induce apoptosis.