海马程序性坏死在心肺转流诱发大鼠认知功能障碍中的作用  

Role of hippocampal necroptosis in cardiopulmonary bypass induced cognitive impairment in rats

作　　者：张琦 石磊[1] 刘扬[1] 陈岩[1] 刘祥[1] 谢永达 李亚南[2] ZHANG Qi;SHI Lei;LIU Yang;CHEN Yan;LIU Xiang;XIE Yongda;LI Yanan(Department of Anesthesiology,Hebei Children s Hospital,Shijiazhuang 050051,China)

机构地区：[1]河北省儿童医院麻醉科,石家庄市050051 [2]河北医科大学第三医院麻醉科

出　　处：《临床麻醉学杂志》2023年第2期173-177,共5页Journal of Clinical Anesthesiology

基　　金：河北省医学科学研究课题(20220712)。

摘　　要：目的探讨海马程序性坏死在心肺转流(CPB)诱发大鼠认知功能障碍中的作用。方法选取健康雄性SD大鼠45只,6月龄,体重400~450 g,采用随机数字表法将大鼠分为三组:对照组(C组)、CPB组和CPB+程序性坏死抑制剂Nec-1组(N组),每组15只。建立CPB模型前,N组腹腔注射Nec-16.25 mg/kg,C组和CPB组腹腔注射等容量生理盐水。CPB组和N组建立无血预充心脏不停跳CPB模型60 min。于CPB结束后2 d采用旷场试验评估自主运动能力。于CPB结束后3 d采用Morris水迷宫实验评估认知功能。水迷宫实验结束后处死大鼠,分离海马组织,采用流式细胞术检测海马神经元程序性坏死数量,采用Western blot法检测p-RIP1、p-RIP3和p-MLKL蛋白含量,通过透射电镜观察海马神经元的超微结构。结果三组旷场试验运动速度、路程及中心停留时间差异无统计学意义。与C组比较,CPB组和N组逃避潜伏期明显延长,穿越原平台次数明显减少,原平台象限停留时间明显缩短,海马神经元程序性坏死率明显升高,p-RIP1、p-RIP3和p-MLKL蛋白含量明显升高(P<0.05),海马神经元细胞器肿胀,溶酶体破裂,部分细胞核发生染色质溶解。与CPB组比较,N组逃避潜伏期明显缩短,穿越原平台次数明显增多,原平台象限停留时间明显延长(P<0.05),海马神经元程序性坏死率明显降低(P<0.05),p-RIP1、p-RIP3和p-MLKL蛋白含量明显降低(P<0.05),海马神经元细胞器肿胀程度、溶酶体破裂程度及细胞核染色质溶解程度明显减轻。结论心肺转流可能通过增加海马程序性坏死程度诱发大鼠认知功能障碍。Objective To evaluate the role of hippocampal necroptosis in cardiopulmonary bypass(CPB)induced cognitive impairment in rats.Methods Forty-five healthy male SD rats,aged 6 months,weighing 400-450 g,were randomly divided into three groups(n=15 in each group):control group(group C),group CPB and CPB+necroptosis inhibitor Nec-1 group(group N).Group N were injected with Nec-16.25 mg/kg intraperitoneally and the other two groups were injected with the same amount of normal saline intraperitoneally.Then,CPB model of beating heart without blood priming for 60 minutes was established in groups CPB and N immediately.The open field test was used to detect the autonomic movement ability of rats on the second day after CPB,and the Morris water maze test was used to detect the cognitive function of rats on the third day after CPB.After the water maze,the rats were sacrificed,the brain was taken and the hippocampal tissue was separated.Then,the hippocampal necroptosis rate was detected using flow cytometry,and the protein expressions of p-RIP1,p-RIP3 and p-MLKL were detected by Western blot,the ultrastructure of hippocampal neurons was observed under transmission electron microscope.Results There were no significant difference in movement speed,distance and central residence time among the three groups.Compared with group C,the escape latency was prolonged,the number of crossing the original platform was reduced,the residence time in the original platform quadrant was shortened,the hippocampal necroptosis rate was increased,the expressions of p-RIP1,p-RIP3 and p-MLKL were up-regulated(P<0.05),the organelles of hippocampal neurons swelled,lysosomes broke,and some chromatin in nuclei dissoluted in group CPB and group N.Compared with group CPB,the escape latency was shortened,the number of crossing the original platform was deduced,the residence time in the original platform quadrant was prolonged,the hippocampal necroptosis rate was decreased,the expressions of p-RIP1,p-RIP3 and p-MLKL were down-regulated,the degree of organelle

关 键 词：海马 程序性坏死 心肺转流 认知功能障碍 

分 类 号：R614[医药卫生—麻醉学]