IP3R1调节的线粒体相关内质网膜结构改变在多次七氟烷麻醉致新生小鼠远期认知功能障碍中的作用  

Role of IP3R1-regulated changes in mitochondria-associated endoplasmic reticulum membrane structure in long-term cognitive impairment induced by multiple exposures to sevoflurane anesthesia in neonatal mice

作　　者：刘春晓[1] 张佳杰 李亚南[1] 石磊[3] 张琦 Liu Chunxiao;Zhang Jiajie;Li Yanan;Shi Lei;Zhang Qi(Department of Anesthesiology,Hebei Children′s Hospital,Shijiazhuang 050051,China;Graduate School of Hebei Medical University,Shijiazhuang 050011,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]河北医科大学研究生学院,石家庄050011 [3]河北省儿童医院麻醉科,石家庄050030

出　　处：《中华麻醉学杂志》2025年第1期59-64,共6页Chinese Journal of Anesthesiology

基　　金：国家自然科学基金(82301382);河北省自然科学基金(H2022316001);河北省政府资助临床医学优秀人才项目(ZF2023126);河北省医学科学研究项目(20240189)。

摘　　要：目的评价1,4,5三磷酸肌醇受体1(IP3R1)调节的线粒体相关内质网膜(MAMs)结构改变在多次七氟烷麻醉致新生小鼠远期认知功能障碍中的作用。方法SPF级健康新生C57BL/6J小鼠60只,雌雄不拘,6日龄,体质量6~10 g,采用随机数字表法分为3组(n=20):对照组(C组)、多次七氟烷麻醉组(S组)和IP3R拮抗剂2-APB+多次七氧烷麻醉组(I+S组)。S组和I+S组于出生后6、8和10 d时吸入3%七氟烷麻醉2 h;I+S组于每次吸入七氟烷麻醉前腹腔注射2-APB 3 mg/kg。于出生后31 d时行旷场实验评估自发活动能力;于出生后31~36 d时行Morris水迷宫实验评估认知功能。水迷宫实验结束后处死小鼠分离海马CA1区,采用流式细胞术检测细胞质游离钙离子浓度([Ca^(2+)]_(i))及程序性坏死率,采用Western blot法检测IP3R1、G蛋白偶联受体75(GRP75)、受体相互作用蛋白激酶1(RIPK1)、RIPK3和磷酸化人混合系列蛋白激酶样结构域(p-MLKL)的表达,透射电镜下观察并记录MAMs长度、内质网周长和线粒体周长。结果3组旷场实验运动速度、运动路程及旷场中心停留时间比较差异无统计学意义(P>0.05)。与C组比较,S组小鼠出生后33~35 d时逃避潜伏期延长,穿越原平台位置次数减少,海马CA1区程序性坏死率和[Ca^(2+)]_(i)升高,IP3R1、GRP75、RIPK1、RIPK3和p-MLKL表达上调,海马神经元MAMs长度/内质网周长比值和MAMs长度/线粒体周长比值升高(P<0.05)。与S组比较,I+S组小鼠出生后32~35 d时逃避潜伏期缩短,穿越原平台位置次数增加,海马CA1区程序性坏死率和[Ca^(2+)]_(i)降低,IP3R1、GRP75、RIPK1、RIPK3和p-MLKL表达下调,海马神经元MAMs长度/内质网周长比值和MAMs长度/线粒体周长比值降低(P<0.05)。结论IP3R1表达上调介导的海马CA1区MAMs结构改变参与了多次七氟烷麻醉致新生小鼠远期认知功能障碍的过程。ObjectiveTo evaluate the role of inositol 1,4,5 triphosphate receptor 1(IP3R1)-regulated changes in mitochondria-associated endoplasmic reticulum membrane(MAM)structure in the long-term cognitive impairment induced by multiple exposures to sevoflurane anesthesia in neonatal mice.MethodsSixty SPF-grade healthy neonatal C57BL/6J mice of either sex,aged 6 days,weighing 6-10 g,were divided into 3 groups(n=20 each)using a random number table method:control group(group C),multiple sevoflurane anesthesia group(group S),and IP3R antagonist 2-APB+multiple sevoflurane anesthesia group(group I+S).Group S and group I+S inhaled 3%sevoflurane anesthesia for 2 h starting from 6,8 and 10 days after birth.In group I+S,2-APB 3 mg/kg was intraperitoneally injected before each sevoflurane anesthesia.The open field test was performed at day 31 after birth to assess the spontaneous mobility.The Morris water maze test was performed at days 31-36 after birth to assess the cognitive function.Mice were sacrificed at the end of the water maze test,hippocampal CA1 region was isolated and hippocampal tissues were obtained for determination of the intracellular calcium ion concentration([Ca^(2+)]_(i))and rate of necroptosis(using Flow cytometry)and expression of IP3R1,G protein-coupled receptor 75(GRP75),receptor-interacting protein kinase 1(RIPK1),RIPK3,and phosphorylated human mixed-series protein kinase-like structural domains(p-MLKL)(by Western blot).Transmission electron microscopy was performed to observe and record the partial length of MAMs,endoplasmic reticulum circumference and mitochondrial circumference.ResultsThere were no statistically significant differences in the speed,distance,and time of staying at the center in open field tests among the three groups(P>0.05).Compared with group C,the escape latency was significantly prolonged on postnatal days 33-35,the number of crossing the original platform was reduced,the necroptosis rate in the hippocampal CA1 region and[Ca^(2+)]_(i) were increased,the expression of IP3R,GRP75,RIPK1,R

关 键 词：七氟醚 认知功能障碍 婴儿 新生 肌醇1 4 5三磷酸受体 线粒体 内质网 偶联 

分 类 号：R614[医药卫生—麻醉学]