NF-кB介导促红细胞生成素神经细胞保护作用的研究

目的:观察促红细胞生成素(EPO)对谷氨酸(Glu)诱导的大脑皮质神经元损伤的保护作用和核因子-кB(NF-кB)介导的细胞内信号转导机制.方法:采用1日龄Wistar大鼠皮层神经细胞体外原代培养技术,建立Glu兴奋性毒性神经细胞损伤模型.实验分为正常对照组、Glu组、EPO组和吡咯烷二硫氨基甲酸(PDTC,NF-кB活化阻断剂)组.倒置显微镜下观察细胞的形态学改变;MTT法测定细胞存活率;流式细胞术测定细胞凋亡率评价细胞损伤程度.结果:Glu处理后神经细胞形态明显受损,EPO组神经细胞形态趋于正常,PDTC组细胞形态与Glu组相似;与正常对照组相比.Glu组神经细胞存活率明显下降、凋亡率明显升高,差异均有统计学意义(P<0.01);而与Glu组相比,EPO组神经细胞存活率明显增高下降、凋亡率明显下降,差异均有统计学意义(P<0.01).与EPO组相比,PDTC组细胞存活率明显下降、凋亡率明显升高,差异均有统计学意义(P<0.01),这些变化与Glu组相似.结论:EPO对Glu诱导的大脑皮质神经元损伤的具有保护作用,其胞内信号转导机制涉及到NF-кB的活化.

NF-кB-mediated protective effect of erythropoitin on neuron against glutamate-induced damage

AIM: To investigate protective effect and possible mechanisms of erythropoitin (EPO) on cortical neuron against damage induced by glutamate(Glu). METHODS: Cortical neurons were removed from 1-day-old newborn rat and then cultured in vitro. The model of damage induced by Glu was established. The experiment was designed as: control group, Glu treatment group,the group of pretreatment with EPO, pyrrolidine dithiocarbamates(PDTC) treatment group. The changes of morphology were observed under inverted microscopy; the cell viability was detected by MTT assay; and the apoptosis rate of neuron was measured by flow cytometry. RESULTS: After exposure to Glu, neurons were obviously damaged. Neurons morph of EPO pretreatment group was similar to that of blank group. However, in PDTC treatment group, cells were obviously damaged, and morph of cells was similar to that of Glu treatment group. Compared with control group, survival of neurons of Glu treatment group significantly decreased, while the neurons apoptotis of Glu treatment group was significantly increased (P<0h01). Compared with Glu treatment group, the neurons, survival of EPO pretreatment group significantly increased, the neurons apoptotis can significantly decrease (P<0h01)ls. Compared with EPO pretreatment group, the cells survival of PDTC treatment group was significantly decreased, while the neurons, apoptotis of Glu treatment group was significantly increased (P<0h01), and similar to that of Glu treatment group. CONCLUSION: EPO could protect cortical neuron from Glu toxicity,which might related to signal transduction of NF-kappaB.