P2X7受体抑制剂腹腔注射对红藻氨酸诱导小鼠癫痫发作及海马损伤的影响  

Effects of P2X7 receptor inhibitor on kainic acid-induced epileptogenesis and hippocampal damage in mice

作　　者：韦彩川 张海菊[1] 叶静萍[1] 姚宝珍[1] WEI Caichuan;ZHANG Haiju;YE Jingping;YAO Baozhen(Department of Pediatrics,Renmin Hospital of Wuhan University,Wuhan 430060,China;不详)

机构地区：[1]武汉大学人民医院儿科,武汉430060

出　　处：《山东医药》2024年第17期7-11,共5页Shandong Medical Journal

基　　金：国家自然科学基金资助项目(81901318)。

摘　　要：目的观察P2X7受体(P2X7R)抑制剂对红藻氨酸诱导小鼠癫痫发作及海马损伤的影响。方法雄性C57BL/6小鼠96只,随机分为正常组36只、模型组36只、P2X7R抑制剂组24只。P2X7R抑制剂组于造模前30 min腹腔注射P2X7R特异性抑制剂A438079,模型组、正常组在同时点注射等量生理盐水。模型组、P2X7R抑制剂组腹腔注射红藻氨酸建立癫痫模型,正常组腹腔注射等量生理盐水。采用脑电图观察小鼠癫痫发作情况,Western blotting法检测小鼠海马组织P2X7R蛋白,尼氏染色观察小鼠海马CA1区损伤情况。采用旷场实验观察正常组、模型组小鼠焦虑样行为,若模型组存在焦虑样行为,则进一步将模型组分为焦虑对照亚组及P2X7R抑制亚组,P2X7R抑制亚组每天腹腔注射A438079,焦虑对照亚组每天腹腔注射等量生理盐水,持续14 d后再次进行旷场实验。采用水迷宫实验观察小鼠认知功能,若模型组存在认知功能损害,则进一步将模型组分为认知功能对照亚组及P2X7R抑制亚组,处理方式同上,持续14 d后再次进行水迷宫实验。结果脑电图记录结果显示,正常组未出现癫痫脑电图波形;模型组出现癫痫持续状态,脑电图表现出强放电功率及高振幅;P2X7R抑制剂组出现癫痫发作,但脑电图放电功率及振幅均较模型组减轻;脑电图总功率及平均振幅模型组>P2X7R抑制剂组>正常组(P均<0.05)。模型组海马组织P2X7R蛋白表达高于正常组,P2X7R抑制剂组海马组织P2X7R蛋白表达低于模型组(P均<0.05)。尼氏染色结果显示,对照组海马CA1区结构完整,存活神经细胞数多,尼氏体较大且较多;模型组海马CA1区结构完整性被破坏,神经细胞数量少,尼氏体数量减少;P2X7R抑制剂组海马CA1区结构、神经细胞及尼氏体数量均较模型组有所改善。旷场实验结果显示,模型组进入中心区域次数及中心区域停留时间均少于正常组,焦虑对照亚组进入中心区域次数及中心Objective To investigate the effects of P2X7 receptor(P2X7R)inhibitor on kainic acid-induced epileptogenesis and hippocampal damage in mice.Methods Ninety-six male C57BL/6 mice were randomly divided into the control group of 36,the model group of 36,and the P2X7R inhibitor group of 24.Mice in the P2X7R inhibitor group were injected intraperitoneally with A438079,a P2X7R-specific inhibitor,30 min before modeling,and mice in the model and control groups were injected with the equal volume of normal saline at the same time point.The epilepsy model was established by intraperitoneal injection of kainic acid in the model group and the P2X7R inhibitor group,and an equal volume of normal saline was injected intraperitoneally in mice of the control group.The severity of seizures was assessed according to the electroencephalography(EEG).Western blotting was used to determine the level of hippocampal P2X7R expression,and Nissl staining was used to assess the extent of hippocampal damage in the CA1 region.Anxiety-like behaviors of mice in the control and model groups were observed in the open-field test.If anxiety-like behaviors existed in the model group,the model group was further divided into the anxiety control subgroup and P2X7R-inhibited subgroup;mice in the P2X7Rinhibited subgroup were injected with A438079 intraperitoneally every day,and mice in the anxiety control subgroup were injected with an equal volume of normal saline every day intraperitoneally,and open-field test was conducted again after 14 days.The morris water maze was used to observe the cognitive function of the mice,and if there was cognitive impairment in the model group,the model group was further divided into the cognitive control subgroup and P2X7R-inhibited subgroup.Mice in the P2X7R-inhibited subgroup were injected intraperitoneally with 15 mg/kg of A438079 every day,while mice in the cognitive control subgroup were injected with the equal volume of normal saline every day,and then the morris water maze was performed again after 14 days.Results

关 键 词：P2X7受体 P2X7受体抑制剂 癫痫 海马损伤 焦虑 

分 类 号：R741.02[医药卫生—神经病学与精神病学]