黄芪甲苷对谷氨酸诱导大鼠海马神经元损伤的保护作用

Protective effects of astragaloside on the damage induced by glutamate in hippocampal neurons

目的:探讨黄芪甲苷对谷氨酸诱导大鼠海马神经元损伤的保护作用及其机制。方法:将培养7~9天大鼠海马细胞随机分为对照组、黄芪甲苷组、谷氨酸损伤组、黄芪甲苷预处理组。采用噻唑蓝法测定细胞存活率,比色法测定培养液中乳酸脱氢酶含量,流式细胞仪测定细胞凋亡率;2,7-二氢二氯荧光黄双乙酸钠探针法检测细胞中活性氧的水平,酶联免疫吸附法检测细胞上清液中8-羟基脱氧鸟苷的含量。结果:0.1 mmol/L谷氨酸作用于海马神经元24小时后,细胞存活率下降、培养液中乳酸脱氢酶含量、细胞凋亡率、细胞中活性氧的水平及上清液中8-羟基脱氧鸟苷的分泌量均明显升高。于损伤前加入黄芪甲苷(0.08,0.40,2.00mg/L)孵育30 min可以不同程度提高细胞存活能力,减少乳酸脱氢酶的漏出,降低细胞凋亡率和上清液中8-羟基脱氧鸟苷的分泌量,但以0.40,2.00 mg/L黄芪甲苷预处理组效果更明显。单纯黄芪甲苷组存活率、细胞凋亡率与正常对照组相比无显著差异,但细胞中活性氧的水平以及上清液中8-羟基脱氧鸟苷的分泌量明显降低。结论:黄芪甲苷预处理能减轻谷氨酸诱导的海马神损伤,其机制可能与其抗氧化有关。

Objective: To investigate the effects of astragaloside on the damage induced by glutamate in hippocampal neurons.Methods: The neurons isolated from hippocampus in new born rats were cultured for 7 ~ 9 days.The cells were randomly divided into control group,astragaloside group,glutamate exposed group and astragaloside protected group.The cell viability was analyzed by MTT assay; the apoptosis rate was measured by flow cytometry.The levels of reactive oxygen species in cells were detected by fluorescent probe DCFH-DA; the level of 8-OHDG was determined by enzyme linked immunosorbent assay.Results: Compared with control group,after exposed of glutamate for 24 h,the viability of neurons were decreased,LDH release and cell apoptosis were increased.While after pretreatment with astragaloside,the rate of surviving neuron were elevated,LDH release,apoptosis ratio,the levels of reactive oxygen species and the contents of 8-OHDG were reduced in different degree.Conclusion: Astragaloside has protective effect on the damage induced by glutamate.The mechanism may be related to antioxidation.