地黄寡糖对谷氨酸诱导海马神经元损伤的影响

The effect of Rehmannia glutinosa oligosaccharides on the damage induced by glutamate in hippocampal neurons

目的观察地黄寡糖对谷氨酸诱导海马神经元损伤的影响。方法将培养7～9d的SD大鼠海马细胞进行NSE鉴定,然后随机分为:正常组(control);单纯地黄寡糖用药组(ROS);谷氨酸损伤组(Glu);谷氨酸损伤前地黄寡糖预处理组(ROS+Glu)。采用倒置显微镜观察细胞形态学变化,噻唑蓝(MTT)法测定细胞存活率,比色法测定培养液中的乳酸脱氢酶(LDH)含量,流式细胞术测定细胞凋亡率。结果0.1mmol·L-1谷氨酸作用于海马神经元24h,出现明显细胞损伤,细胞存活率下降,培养液中LDH含量明显增加,细胞凋亡率增高(P<0.01);4、20、100mg·L-1的地黄寡糖可不同程度的改善谷氨酸损伤引起的神经细胞形态的改变,提高细胞存活力,减少LDH的漏出,降低细胞凋亡率,并呈一定的剂量依赖性。4、20、100mg·L-1单纯地黄寡糖组与正常组间各指标差异没有显著性(P>0.05)。结论谷氨酸能诱导海马原代细胞的凋亡,地黄寡糖能有效保护海马神经细胞免受谷氨酸诱导的细胞损伤。

Aim To investigate the effects of Rehmannia glutinosa oligosaccharides （ROS） on the damage induced by glutamate in hippocampal neurons. Methods The neurons isolated from hippocampus in new born SD rats were cultured for 7 - 9 days, which were specifically stained with NSE and then randomly divided into four groups ： （ Ⅰ ） Normal cultures （control） ; （ Ⅱ ） ROS control cultures; （ Ⅲ ） Glutamate-exposed control cultures; （Ⅳ） Glutamate-exposed cultures pretreated with ROS. The neurons morphology was observed under inverted microscope; cell viability was assayed by MTT staining; LDH release was detected with chromatometry and flow cytometric analysis for identification and quantification of cell apoptosis. Results Compared with normal group, after exposure of glutamate for 24 h, the viability of neurons was decreased, LDH release and cell apoptosis were increased （P 〈 0. 01 ）. While after pretreatment with ROS, the rate of surviving neuron was elevated, LDH release and apoptosis ratio were reduced in different degree, depending on dose in certain range. Conclusions Glutamate can induce the apoptosis of hippocampus neurons, and ROS can protect neurons the damage induced by glutamate effectively.