刺五加皂甙对皮质神经元缺血缺氧性损伤的保护作用

Protective effects of Acanthopanax senticousus saponins on cortical neuronal ischemia-hypoxia injury

目的探讨刺五加皂甙(ASS)对皮质神经元缺血缺氧性损伤的保护作用。方法胎鼠皮质神经元经缺血缺氧和谷氨酸(Glu)作用模拟缺血性皮质神经元损伤模型。随机分成缺血缺氧组(HI组)、Glu组、ASS组和对照组;ASS组在实验前、中均加入终浓度为50mg/L的ASS。用流式细胞仪检测神经元凋亡率,用硝酸还原酶法测定细胞培养上清液中一氧化氮(NO)的含量,用3[4,5二甲基磺胺噻唑]2,5二苯基四唑溴化物(MTT)比色法、乳酸脱氢酶(LDH)释放量测定神经元活性,电镜下观察细胞形态学变化。结果(1)随着缺血缺氧时间延长神经元存活率逐渐下降,至缺血缺氧8h达到高峰;Glu作用的神经元,其存活率随作用时间延长而下降。(2)HI组及Glu组皮质神经元存活率显著低于正常对照组,LDH释放量、NO含量及凋亡率均显著高于对照组(均P<0.05);与HI组和Glu组比较,HI+ASS组及Glu+ASS组皮质神经元的存活率显著升高,而LDH释放量、NO含量及凋亡率均显著降低(均P<0.05)。结论ASS能提高缺血神经元存活率、降低凋亡率,抑制NO和LDH释放,从而起到神经保护作用。

Objective To explore the protective effects of Acanthopanax senticousus saponins （ASS） on ischemia-hypoxia injury of cortical neuron. Methods The models of cortical neuron damage induced by hypoxiaischemia （HI） and glutamate （Glu） were established on cultured embryonic cortical neurons. The neurons were randomly divided into HI group, Glu group, ASS group and control group. ASS （50 mg/L） was added into the ASS group before and during neuron damage. The rate of neuronal apoptosis was measured by flow cytometer, nitric oxide （NO） content was determined by Nitrate reductive assay and neuron survival was measured by MTF assay and the release of LDH. Morphologic change of neurons was observed under electron microscopy. Results （ 1 ） The cortical neuron survival decreased time-dependently in HI group and reached peak at 8h after hypoxia-ischemia. Glutamate leaded the cortical neuron survival decreasing time-dependently. （2） Compared with the control group, the cortical neuron survival decreased in HI group and Glu group, but the neuron apoptosis, the release of LDH and NO contents increased significantly （ all P 〈 0. 05 ）. The neuron survival in ASS group was significantly higher than those in HI group and Glu group, but the neuron apoptosis, release of LDH and NO contents decreased （all P〈 0.05 ）. Conclusions ASS may play neuroprotectvie role by increasing neuron survival, decreasing neuron apoptosis, and inhibiting the release of NO and LDH.