雷公藤单体T10对谷氨酸所致PC12细胞损伤的保护作用及机制研究

目的:以谷氨酸为工具药建立帕金森病(Parkinson disease, PD)的氧化应激模型,观察雷公藤内酯醇(triptolide, T10)的保护作用并初步探讨其作用机制.方法:分别用10-10 、10-9 mol*L-1的T10和/或10、50 mmol*L-1谷氨酸处理PC12细胞24 h,MTT法检测细胞的活力,并用AnnexinV-FITC和PI对活细胞进行双标,流式细胞仪检测凋亡细胞百分率.分别采用荧光探剂2′7′-二氯荧光素乙酰乙酸盐和JC1定量检测细胞内活性氧和线粒体膜电位水平以探讨其作用机制.结果:谷氨酸处理PC12细胞24 h,可引起细胞的凋亡和坏死,细胞内活性氧形成明显增加,线粒体膜电位下降.而用10-10或10-9 mol*L-1的T10预保护30 min可明显减轻PC12细胞的损伤,并抑制细胞内活性氧的形成和线粒体膜电位下降.结论:10-10或10-9 mol*L-1的T10能有效地拮抗谷氨酸对PC12细胞的损伤作用.其作用机制可能与抑制谷氨酸诱导的细胞内活性氧自由基的生成和线粒体膜电位下降有关.

Neuroprotective effects of Tripterygium Wilforddi Hook F monomer T10 on glutamate induced PC12 cell line damage and its mechanism

OBJECTIVE: To study the neuroprotective effect and related mechanisms of triptolide. METHODS: After being treated with 10(-10), 10(-9) mol.L-1 triptolide and/or 10, 50 mmol.L-1 glutamate for 24 hours, and the viability of PC12 was detected by MTT conversion, and the apoptosis rate was detected by AnnexinV-FITC and PI staining. In order to understand the underlying mechanism of the neuroprotective effect of triptolide, H2DCFDA and JC1 were used to detect the reactive oxygen species (ROS) and mitochondrial membrane potential. RESULTS: Glutamate could induce PC12 necrosis and apoptosis. T10 at 10(-10) or 10(-9) mol.L-1 inhibited glutamate-induced cell death, ROS formation and decrease of mitochondrial membrane potential. CONCLUSION: T10 at 10(-10) or 10(-9) mol.L-1 can protect PC12 from the death induced by glutamate; and the underlying mechanisms may be involved in the inhibition of ROS formation and decrease of mitochondrial membrane potential.