IP3受体在七氟烷麻醉致老龄大鼠海马神经元程序性坏死中的作用  

Role of IP3 receptor in necroptosis of hippocampal neurons induced by sevoflurane anesthesia in aged rats

作　　者：李伟[1] 张琦 于家旭 王旭鹏 尹春平[1] 周琦 高明阳 王秋筠[1] Li Wei;Zhang Qi;Yu Jiaxu;Wang Xupeng;Yin Chunping;Zhou Qi;Gao Mingyang;Wang Qunjun(Department of Anesthesiology,Third Hospital of Hebei Medical University,Shijiazhuang 050051,China;Department of Anesthesiology,Hebei Children′s Hospital,Shijiazhuang 050030,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

出　　处：《中华麻醉学杂志》2022年第12期1456-1460,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1,4,5三磷酸肌醇受体(IP3R)在七氟烷麻醉致老龄大鼠海马神经元程序性坏死中的作用。方法:健康雄性SD大鼠60只,18月龄,体重500~600 g,采用随机数字表法分为3组(n=20):对照组(C组)、七氟烷麻醉组(S组)和七氟烷麻醉+IP3R拮抗剂组(S+I组)。S组和S+I组吸入2%七氟烷5 h。S+I组吸入七氟烷前10 min时腹腔注射IP3R拮抗剂2-APB 3 mg/kg,C组和S组腹腔注射同容量二甲基亚砜。七氟烷麻醉结束后当天采用Morris水迷宫实验检测认知功能,然后处死大鼠取脑组织,行HE染色和尼氏染色,光镜下观察病理学结果;采用流式细胞术检测海马神经元胞浆游离钙离子浓度([Ca^(2+)]i)和程序性坏死率;采用Western blot法检测IP3R、受体相互作用蛋白1(RIPK1)、受体相互作用蛋白3(RIPK3)和磷酸化混合系列蛋白激酶样结构域(p-MLKL)的表达。结果:与C组比较,S组和S+I组逃避潜伏期延长,穿越平台次数减少,目标象限停留时间缩短,海马神经元[Ca^(2+)]i和程序性坏死率升高,海马神经元IP3R、RIPK1、RIPK3和p-MLKL表达上调(P<0.05);与S组比较,S+I组逃避潜伏期缩短,穿越平台次数增多,目标象限停留时间延长,海马神经元[Ca^(2+)]i和程序性坏死率降低,海马神经元IP3R、RIPK1、RIPK3和p-MLKL表达下调(P<0.05)。结论:七氟烷麻醉致老龄大鼠认知功能障碍的机制可能与IP3R激活介致钙稳态失衡,从而诱发程序性坏死有关。To evaluate the role of 1,4,5-inositol triphosphate receptor(IP3R)in necroptosis of hippocampal neurons induced by sevoflurane anesthesia in aged rats.Methods Sixty healthy male Sprague-Dawley rats,aged 18 months,weighing 500-600 g,were divided into 3 groups(n=20 each)using a random number table method:control group(group C),sevoflurane anesthesia group(group S)and sevoflurane anesthesia+IP3R antagonist group(group S+I).S and S+I groups inhaled 2%sevoflurane for 5 h.In group S+I,IP3 receptor antagonist 2-APB 3 mg/kg was intraperitoneally injected at 10 min before sevoflurane inhalation,and the equal volume of dimethyl sulfoxide was intraperitoneally injected in group C and group S.Morris water maze test was used to test the cognitive function on the day after the end of sevoflurane anesthesia.Then the animals were sacrificed and the brain tissues were obtained for microscopic examination of the pathological changes after HE staining and Nissl staining(with a light microscope)and for determination of the free calcium concentration([Ca^(2+)]i)and rate of necroptosis of hippocampal neurons(by flow cytometry)and expression of IP3R,receptor-interacting protein kinase-1(RIPK1),receptor-interacting protein kinase-3(RIPK3)and phosphorylated mixed lineage kinase domain-like protein(p-MLKL)(by Western blot).Results Compared with group C,the escape latency was significantly prolonged,the times of crossing the platform were reduced,the time of staying at the target quadrant was shortened,the[Ca^(2+)]i and necroptosis rate of hippocampal neurons were increased,and the expression of IP3R,RIPK1,RIPK3 and p-MLKL in hippocampal neurons was up-regulated in group S and group S+I(P<0.05).Compared with group S,the escape latency was significantly shortened,the times of crossing the platform were increased,the time of staying at the target quadrant was prolonged,the[Ca^(2+)]i and necroptosis rate of hippocampal neurons were decreased,and the expression of IP3R,RIPK1,RIPK3 and p-MLKL in hippocampal neurons was down-regulated in group S

关 键 词：肌醇1 4 5-三磷酸受体 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]