NMDA受体在七氟烷麻醉致老龄小鼠海马神经元程序性坏死中的作用  

Role of NMDA receptors in sevoflurane anesthesia-caused necroptosis in hippocampal neurons of aged mice

作　　者：于家旭 张琦 尹春平[1] 李亚南[1] 李伟[1] 朱炼[3] 侯志勇[3] 王秋筠[1] Yu Jiaxu;Zhang Qi;Yin Chunping;Li Yanan;Li Wi;Zhu Lian;Hou Zhiyong;Wang Qiujun(Department of Anesthesiology,Third Hospital of Hebei Medical University,Shijiazhuang 050051,China;Department of Anesthesiology,Hebei Children′s Hospital,Shijiazhuang 050030,China;Center of Emergency and Trauma,Third Hospital of Hebei Medical University,Shijiazhuang 050051,China)

机构地区：[1]河北医科大学第三医院麻醉科,石家庄050051 [2]河北省儿童医院麻醉科,石家庄050030 [3]河北医科大学第三医院创伤与急救中心,石家庄050051

出　　处：《中华麻醉学杂志》2022年第1期55-59,共5页Chinese Journal of Anesthesiology

基　　金：国家自然科学基金面上项目(81771134);河北省自然科学基金面上项目(H2018206305)。

摘　　要：目的评价N-甲基-D-天冬氨酸受体(NMDA受体)在七氟烷麻醉致老龄小鼠海马神经元程序性坏死中的作用。方法清洁级健康雄性C57BL/6小鼠90只,18月龄,体重27~30 g,采用随机数字表法分为3组(n=30):对照组(C组)、七氟烷麻醉组(S组)和七氟烷麻醉+NMDA受体拮抗剂盐酸美金刚组(S+M组)。S组和S+M组小鼠连续3 d吸入3%七氟烷2 h,S+M组于每次吸入七氟烷前1 h腹腔注射盐酸美金刚20 mg/kg,C组只吸入纯氧。分别于麻醉前1 d、麻醉后3和7 d时每组随机取10只小鼠行Morris水迷宫实验。Morris水迷宫实验结束后立即处死小鼠取海马,于光镜下观察病理学结果,采用流式细胞术测定神经元程序性坏死率和胞浆游离钙离子浓度([Ca^(2+)]i),Wes-tern blot法检测NMDA受体亚型GluN2A、GluN2B和受体相互作用蛋白激酶1(RIP1)的表达。结果与C组比较,S组和S+M组麻醉后各时点逃避潜伏期延长,穿越原平台位置次数减少,海马[Ca^(2+)]i和神经元程序性坏死率升高,GluN2A、GluN2B和RIP1表达上调(P<0.05),病理学损伤加重;与S组比较,S+M组麻醉后各时点逃避潜伏期缩短,穿越原平台位置次数增加,海马[Ca^(2+)]i和神经元程序性坏死率降低,GluN2A、GluN2B和RIP1表达下调(P<0.05),病理学损伤减轻。结论NMDA受体参与了七氟烷麻醉致老龄小鼠认知功能障碍的过程,其机制可能与促进海马神经元程序性坏死有关。Objective To evaluate the role of N-methyl-D-aspartate receptors(NMDA receptors)in sevoflurane anesthesia-caused necroptosis in hippocampal neurons of aged mice.Methods Ninety clean-grade healthy male C57BL/6 mice,aged 18 months,weighing 27-30 g,were divided into 3 groups(n=30 each)using a random number table method:control group(group C),sevoflurane anesthesia group(group S)and sevoflurane anesthesia plus NMDA receptor antagonist memantine hydrochloride group(group S+M).Mice inhaled 3%sevoflurane for 2 h for 3 consecutive days in S group and S+M group,and memantine hydrochloride 20 mg/kg was intraperitoneally injected at 1 h before each inhalation of sevoflurane in S+M group.Mice only inhaled pure oxygen for 2 h in group C.Ten mice of each group were selected on 1 day before anesthesia and 3 and 7 days after anesthesia to perform Morris water maze test.The mice were sacrificed immediately after Morris water maze test,and hippocampus was removed for microscopic examination of pathological changes(with a light microscope)and for determination of the necroptosis rate of neurons and cytoplasmic free calcium concentration([Ca^(2+)]i)(by flow cytometry),and expression of NMDA receptor subtypes GluN2A,GluN2B and receptor-interacting protein kinase 1(RIP1)(by Western blot).Results Compared with group C,the escape latency was significantly prolonged,and the frequency of crossing the original platform was decreased,and the[Ca^(2+)]i and neuronal necroptosis rate in the hippocampus were increased at each time point after anesthesia,and the expression of GluN2A,GluN2B and RIP1 was up-regulated(P<0.05),and the pathologic changes were accentuated in S group and S+M group.Compared with group S,the escape latency was significantly shortened,and the frequency of crossing the original platform was increased,and the[Ca^(2+)]i and neuronal necroptosis rate in the hippocampus were decreased at each time point after anesthesia,and the expression of GluN2A,GluN2B and RIP1 was down-regulated(P<0.05),and the pathologic changes were atte

关 键 词：受体 N-甲基-D-天冬氨酸 麻醉药 吸入 老年人 海马 神经元 坏死 

分 类 号：R614[医药卫生—麻醉学]