摘要目的探讨癫(痫)持续状态(SE)大鼠模型中海马神经元损伤、谷氨酸α-氨基-3-羟基-5-甲基异(噁)唑-4-丙酸(AMPA)受体第二亚单位GluR2的表达变化以及是否存在GluR2与甘油醛-3-磷酸脱氢酶(GAPDH)蛋白复合物的耦合变化.方法 应用氯化锂-毛果芸香碱建立SE大鼠模型(62只),同时设立正常对照组(20只),按照随机数字表法将建SE大鼠分为SE后1h组(6只)、6h组(12只)、24h组(12只)、72 h组(12只)及7d组(20只).按照随机数字表法选择正常对照及SE后6h、24 h、72 h、7d(各6只)作Nissl染色、原位末端标记(TUNEL)分别观察大鼠海马形态学变化、凋亡发生;再选择对照组及SE后1h、6h、24 h、72 h及7d组大鼠(各6只)用Western blot检测GluR2蛋白的表达变化,用免疫共沉淀及Western blot技术研究GluR2与GAPDH蛋白复合物的耦合变化情况.结果 SE后各时间点海马CA1、CA3区神经细胞数量显著减少,与正常对照组比较差异有统计学意义(F=30.866、24.043,P均＜0.05);SE后24h、72 h、7d海马CA1、CA3区凋亡细胞数量增加,与正常对照组比较差异有统计学意义(F=84.762、52.574,P均＜0.01);与正常对照组比较,SE后1h、6h海马GluR2蛋白相对表达量减少,但差异无统计学意义(P＞0.05),SE后24 h、72 h、7 d GluR2相对表达量减少,差异有统计学意义(F=76.506,P＜0.01);SE后72 h与正常对照组比较,GluR2与GAPDH形成的蛋白复合物耦合增加,差异有统计学意义(t=7.029,P＜0.05).结论 SE可导致海马区神经元损伤,GluR2表达降低及GluR2/GAPDH蛋白复合物耦合增加可能是其机制之一.

abstractsObjective To investigate hippocampal neuronal damage and dynamic change of α-amino-3-hydroxy-5-methyl-4-isoxazole propionic acid (AMPA) receptor subunit GluR2 in status epilepticus, to find out whether GluR2/glyceral dehyde-3-phosphate dehydrogenase (GAPDH) interaction has any change.Methods Male Wistar rats (62 cases) were induced to status epilepticus by using LiC1-pilocarpine.The 62 rats were divided into 1 h (6 cases),6 h (12 cases), 24 h (12 cases),72 h (12 cases)and 7 d (20 cases)after status epilepticus.At the same time, the healthy control group (12 cases) was established.Morphologic changes of hippocampus and the amount of apoptotic cells in healthy control and SE model groups at different time points (6 h, 24 h, 72 h, 7 d) (6 cases each group) after status epilepticus were quantified by adopting Nissl staining and terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end-labeling staining respectively.Expressions of GluR2 in healthy control and SE model groups at 1 h,6 h,24 h,72 h and 7 d (6 cases each group) after status epilepticus were detected by using Western blot.Co-precipitation and Western blot techniques were used to investigate whether the GluR2/GAPDH interaction in the hippocampus was increased.Results Compared with the healthy control group, the number of nerve cells in the hippocampal CA1 and CA3 regions was significantly reduced at all studied time points(F =30.866,24.043, all P ＜0.05).Apoptotic cells in hippocampal CA1 and CA3 regions were significantly increased at 24 h,72 h and 7 d after status epilepticus (F =84.762,52.574, all P ＜ 0.01).GluR2 at 1 h,6 h after status epilepticus was equal to that of the control group (P ＞ 0.05), but it was shown to be significantly down-regulated at other studied time points (F =76.506,P ＜ 0.01);when compared with the healthy control group,the GluR2/GAPDH interaction was significantly up-regulated in the hippocampus at 72 h after status epilepticus (t =7.029, P ＜ 0.05).Conclusions Status epilepticus can lead to neuronal damage in the hippocampus.Down-regulation of GluR2 and increase of the GluR2/GAPDH complex formation might be one of the mechanisms involved in hippocampal neuronal damage.