花生四烯酸乙醇胺诱导人U251胶质瘤细胞增殖凋亡的研究

The effect of anandamide on inhibiting proliferation and inducing apoptosis of human U251 glioma cell line

目的 观察大麻受体激动剂花生四烯酸乙醇胺（AEA）诱导人U251胶质瘤细胞凋亡中的作用.方法 体外培养人U251胶质瘤细胞,并将U251胶质瘤细胞株分为空白对照组,分别含AEA药液1、5、10 μmol/L DMEM培养液共4组,以24、48 h作为药物作用时间进行观察.在倒置显微镜下观察细胞形态变化,通过流式细胞仪检测细胞凋亡,激光共聚焦显微镜观察经Hoechst 33258染色的药物作用细胞爬片.结果 各对照组细胞生长良好,各实验组细胞出现不同程度的细胞变圆、漂浮和细胞碎片;流式细胞仪检测显示通过5 μmol/L AEA作用24 h后,可出现早期细胞凋亡,10 μmol/L AEA作用48 h后早期凋亡率为（43.78±3.45）％;激光共聚焦显微镜证明AEA处理U251细胞后,能诱导细胞凋亡,染色质固缩.结论 AEA可以诱导人U251胶质瘤细胞早期凋亡,并呈剂量、时间依赖性.

Objective To investigate the effects of the early apoptosis induction in U251 glioma cells by specific endogenous CB1 agonist anandamide （AEA） treatment.Methods Human U251 glioma cell line were cultured with DMEM＇ s medium in vitro.Cultured cells were divided into 4 groups：control group,1,5 and 10 μmol/L AEA groups.At the same time,the time line was divided into 2 groups （24 h and 48 h）.The cell morphological changes were observed by invert microscope.The flow cytometer evaluated U251 cells early apoptosis which was induced by AEA.The laser confocal microscopy made further detection to the cell apoptosis.Results The cell morphological changes included cell flotation and fragment.The apoptosis rate investigated by flow cytometer was 43.78％ at 10 μmol/L AEA for 48 h.Cell nucleus pyknosis stained by Hoechst 33258 can by observed by the confocal microscopy.Conclusion AEA could produce apoptosis effect on human U251 glioma cell line in dose-and time-dependent manner.