异氟醚通过激活PERK/eIF2α通路诱导神经元凋亡和ERS在PND中的作用机制  

The mechanism of isoflurane-induced neuronal apoptosis and endoplasmic reticulum stress in perioperative neurocognitive disorders via RNA-activated protein kinase-like endoplasmic reticulum kinase/eukaryotic translation initiation factor 2αpathway

作　　者：董营 何二涛 师荣荣 Dong Ying;He Ertao;Shi Rongrong(Department of Anesthesiology,Dingzhou People's Hospial,Dingzhou 073000,China;Department of Anesthesiology,the Second Hospital of Lanzhou University,Lanzhou730000,China)

机构地区：[1]定州市人民医院麻醉科,定州073000 [2]兰州大学第二医院麻醉科,兰州730000

出　　处：《国际麻醉学与复苏杂志》2022年第8期789-794,共6页International Journal of Anesthesiology and Resuscitation

基　　金：河北省医学科学研究重点课题计划(20181474)。

摘　　要：目的探究异氟醚通过激活蛋白激酶样内质网激酶(protein kinase-like endoplasmic reticulum kinase,PERK)/真核翻译起始因子2α(eukaryotic translation initiation factor 2α,eIF2α)通路诱导神经元凋亡和内质网应激(endoplasmic reticulum stress,ERS)在围手术期神经认知障碍(perioperative neurocognitive disorders,PND)中的作用机制.方法30只雄性SD大鼠按随机数字表法分为3组(每组10只):对照组、异氟醚组和PERK抑制剂组(GSK组).异氟醚组和GSK组大鼠放置于麻醉箱,通过吸入2%异氟醚4h建立PND模型,GSK组大鼠在吸入异氟醚6h前使用PERK抑制剂GSK2606414(150 mg/kg)灌胃;对照组大鼠进行同样处理,但不吸入异氟醚,给予等量生理盐水灌胃.吸入异氟醚4 h后进行水迷宫试验,记录逃避潜伏期、穿越平台次数和目标象限停留时间,分析大鼠神经功能.水迷宫实验结束后处死大鼠取海马组织,TUNEL染色检测海马组织神经元细胞凋亡率,Western blot法检测海马组织PERK、eIF2α磷酸化水平及78kDa葡糖调节蛋白(78kDa glucose-regulated pro-tein,GRP78)、C/EBP同源蛋白(C/EBP homologous protein,CHOP)水平,免疫组化染色检测海马组织B淋巴细胞瘤-2(B-cell lymphoma-2,Bcl-2)、B淋巴细胞瘤-2-Associated X(B-cell lymphoma-2-Associated X,Bax)水平.结果异氟醚组逃避潜伏期长于GSK组及对照组(P<0.05),穿越平台次数和目标象限停留时间少于GSK组及对照组(P<0.05).异氟醚组海马组织神经元细胞凋亡率高于GSK组及对照组(P<0.05),PERK、eIF2α磷酸化水平高于GSK组及对照组(P<0.05),GRP78、CHOP水平高于GSK组及对照组(P<0.05),Bax水平高于GSK组及对照组,Bcl-2水平低于GSK组及对照组(P<0.05).结论异氟醚可能通过激活PERK/eIF2α通路和ERS引起海马组织神经元细胞凋亡,抑制PERK/eIF2α通路的激活可通过抑制ERS缓解海马组织神经元细胞凋亡从而对PND模型大鼠的神经功能起到保护作用.Objective To explore the mechanism of isoflurane-induced neuronal apoptosis and endoplasmic reticulum stress(ERS)through protein kinase-like endoplasmic reticulum kinase(PERK)eukaryotic translation initiation factor 2α(eIF2α)pathway in perioperative neurocognitive disorders(PND).Methods Thirty male Sprague-Dawley(SD)rats were divided into three groups by random number table method with ten rats in each group:the control group,isoflurane group,and PERK inhibitor group(GSK group).Rats in the isoflurane group and GSK group were placed in an anesthesia box,and the PND model was established by inhaling 2%isoflurane for 4 h.The rats in the GSK group were given PERK inhibitor GSK2606414(150 mg/kg)by gavage once 6 h before inhalation of isoflurane.Rats in the control group received the same treatment,but did not inhale isoflurane,and were given the same amount of normal saline by gavage.Four hours after inhalation of isoflurane,the neurological function of the rats was tested by a water maze,the escape latency,the number of mice crossing the platform,and the residence time in the target quadrant were recorded.Afer the experiment,the rats were killed and their hippocampal tssue was taken.TUNEL staining was used to detect the apoptosis rate in the hippocampus.Western blot was used to detect the the phosphorylated levels of PERK and elF2αand the levels of 78 kDa glucose-regulated protein(GRP78)and C/EBP homologous protein(CHOP).Immunohistochemical staining was used to detect B-cell lymphoma-2-Associated X(Bax)and B-cel lymphoma-2(Bel-2)levels in the hippocampus.Results The escape latency in the isoflurane group was higher than that in the GSK group and the control group(P<0.05).The crossed platform times and the target quadrant stay time in the isoflurane group were lower than those in the GSK group and the control group(P<0.05).The phosphorylated levels of PERK and elF2αin the hippocampus of isoflurane group were higher than those of GSK group and control group(P<0.05).The levels of GRP78 and CHOP in the hippocampus of

关 键 词：麻醉 异氟醚 围手术期神经认知障碍 海马 内质网应激 凋亡 

分 类 号：R614[医药卫生—麻醉学]