拮抗P2X7受体在减轻子痫抽搐大鼠海马损伤中的作用  

Role of antagonistic P2X7 receptor in reducing hippocampal damage in epileptic rats

作　　者：李晓晶 李晓兰 Li Xiaojing;Li Xiaolan(Dept of Obstetrics and Gynecology,The First Affiliated Hospital of Anhui Medical University,Hefei 230022;NHC Key Laboratory of Study on Abnormal Gametes and Reproductive Tract,Hefei 230022)

机构地区：[1]安徽医科大学第一附属医院妇产科,合肥230022 [2]国家卫生健康委配子及生殖道异常研究重点实验室,合肥230022

出　　处：《安徽医科大学学报》2024年第10期1736-1741,共6页Acta Universitatis Medicinalis Anhui

基　　金：国家自然科学基金项目(编号:81801477)。

摘　　要：目的研究干预嘌呤能离子通道型受体7(P2X7)受体可减轻子痫抽搐海马损伤的作用机制。方法SD雌鼠28只按随机数字表法分为4组(每组n=7),将雌雄鼠进行2∶1合笼,第2天阴道涂片镜下观察到精子细胞记为妊娠第0天(GD0),通过GD14尾静脉注射LPS、GD16~18连续腹腔注射PTZ的方式制备子痫抽搐孕鼠模型。于GD18腹腔注射PTZ后立即腹腔注射P2X7抑制剂(A-438079)制备抑制剂组。即分组为:①正常妊娠组(P组);②类子痫抽搐组(LPS+PTZ组);③P2X7抑制剂组(LPS+PTZ+A-438079组);④类子痫对照组(LPS+PTZ+NS组)。根据Racine评分标准判断大鼠癫痫发作时等级以此评估各组孕鼠抽搐程度记录各期持续时间;Western blot检测孕鼠海马P2X7、NLRP3蛋白表达水平;ELISA测定孕鼠血清中肿瘤坏死因子-α(TNF-α)、白细胞介素(IL)-1β、胎盘生长因子(PIGF)、可溶性fms样酪氨酸激酶1(sFlt-1)炎症因子表达情况。结果LPS+PTZ组、LPS+PTZ+NS组大鼠与P组大鼠相比,抽搐行为学评分显著增高;LPS+PTZ组、LPS+PTZ+NS组大鼠与LPS+PTZ+A-438079组大鼠相比,1期癫痫发作潜伏期、5期癫痫发作潜伏期、1-4期癫痫发作持续时间减少;5期癫痫发作比率和5期癫痫发作持续时间均延长。Western blot结果显示,与LPS+PTZ组和LPS+PTZ+NS组相比,LPS+PTZ+A-438079组孕鼠海马P2X7、NLRP3表达升高。ELISA结果显示,与P组相比,LPS+PTZ组、LPS+PTZ+NS组孕鼠血清炎症因子标志物TNF-α和IL-1β;先兆子痫标志物sFlt-1;PIGF升高,而给予P2X7受体抑制剂后,各项指标均有所降低。结论干预P2X7受体可能通过降低类子痫抽搐孕鼠的全身炎症反应,减轻神经海马损伤,提高抽搐阈值,抑制子痫抽搐的发生。Objective To investigate the mechanism of purinergic ion channel receptor 7(P2X7)intervention in reducing hippocampal damage in epileptic convulsions.Methods A total of 28 SD female rats were divided into 4 groups(n=7 in each group)according to random number table method.The male and female rats were combined with 2∶1 cage.The sperm cells observed by vaginal smear on day 2 were marked as day 0 of pregnancy(GD0),and the model of epileptic pregnant mice was prepared by injecting LPS into the tail vein of GD14 and PTZ into the abdomen continuously on GD16-18.The inhibitor group was prepared by intraperitoneal injection of P2X7 inhibitor(A-438079)immediately after intraperitoneal injection of PTZ in GD18.They were divided into:①normal pregnancy group(P group);②Epilepsy group(LPS+PTZ group);③P2X7 inhibitor group(LPS+PTZ+A-438079 group);④Eclampsia control group(LPS+PTZ+NS group).Racine scoring criteria were used to determine the grade of epileptic seizure in rats to evaluate the degree of epileptic seizure in each group and record the duration of each phase.The expression levels of P2X7 and NLRP3 in the hippocampus of pregnant mice were detected by Western blot.The expression of TNF-α,IL-1β,PIGF and sFlt-1 inflammatory factors in serum of pregnant mice was determined by ELISA.Results The tic behavioral scores of rats in LPS+PTZ group and LPS+PTZ+NS group were significantly higher than those in P group.Compared with LPS+PTZ+A-438079 group,LPS+PTZ+NS group and LPS+PTZ+NS group,the first stage of seizure latency,the fifth stage of seizure latency and the duration of 1-4 stages of seizure decreased.Both the rate of stage 5 seizures and the duration of stage 5 seizures were prolonged.Western blot results showed that compared with LPS+PTZ group and LPS+PTZ+NS group,the expressions of P2X7 and NLRP3 in hippocampus of pregnant rats in LPS+PTZ+A-438079 group increased.ELISA results showed that compared with P group,serum inflammatory factor markers tumor necrosis factor-α(TNF-α)and interleukin(IL)-1βwere detected

关 键 词：P2X7受体 子痫抽搐 海马损伤 炎症 炎症因子 

分 类 号：R714.7[医药卫生—妇产科学]