缺氧预处理对异丙酚诱导新生大鼠大脑神经毒性的影响  

Effects of hypoxic preconditioning on neurotoxicity induced by propofol in neonatal rats

作　　者：肖飞[1] 梁羽冰[2] 吕靖[1] 关睿聪 谢玉波[1] 利莉[1] Xiao Fei;Liang Yubing;Lyu Jing;Guan Ruicong;Xie Yubo;Li Li(Department of Anesthesiology,the First Affiliated Hospital of Guangxi Medical University,Nanning 530021,China;Department of Anes⁃thesiology,the Affiliated Tumor Hospital of Guangxi Medical University,Nanning 530021,China)

机构地区：[1]广西医科大学第一附属医院麻醉科,南宁530021 [2]广西医科大学附属肿瘤医院麻醉科,南宁530021

出　　处：《国际麻醉学与复苏杂志》2020年第5期422-426,共5页International Journal of Anesthesiology and Resuscitation

基　　金：国家自然科学基金(81560500);广西壮族自治区自然科学基金(2017GXNSFBA198108);广西壮族自治区重点研发计划(桂科AB18221031)。

摘　　要：目的探讨缺氧预处理对异丙酚诱导新生大鼠大脑神经毒性的影响。方法SPF级SD大鼠75只,日龄7 d,体重10~15 g,采用随机数字表法将其分为5组(每组15只):生理盐水组(N组),腹腔注射生理盐水100μl;脂肪乳剂组(F组),腹腔注射脂肪乳剂100μl;异丙酚组(P组),腹腔注射异丙酚100 mg/kg;缺氧预处理组(H组),大鼠置于8%氧浓度10 min,空气10 min,循环5次,空气中恢复2 h;缺氧预处理+异丙酚组(H+P组),先给予缺氧预处理(同H组),后给予异丙酚(同P组)。大鼠苏醒后时点,断头取海马组织:制作电镜标本,应用透射电镜观察各组海马神经细胞的超微结构;制作石蜡切片,行TUNEL法染色,检测TUNEL阳性细胞的数量;Western blot法检测活化的半胱氨酸蛋白酶-3(cleaved-caspase-3)、B淋巴细胞瘤-2(B-cell lymphoma-2,Bcl-2)和B淋巴细胞瘤-2相关X蛋白(B-cell lymphoma-2-associated X protein,Bax)蛋白水平。结果与N组比较,P组海马神经细胞萎缩、细胞器溶解、核仁消失,TUNEL阳性细胞数量明显增加(P<0.05),Bax蛋白和cleaved-caspase-3蛋白水平升高(P<0.05),Bcl-2蛋白水平降低(P<0.05)。与P组比较,H+P组海马神经细胞的细胞膜完整、细胞器和细胞核清晰可见、核染色质稍有减少,TUNEL阳性细胞数量减少(P<0.05),Bax蛋白和cleaved-caspase-3蛋白水平降低(P<0.05),Bcl-2蛋白水平升高(P<0.05)。结论缺氧预处理可以减轻异丙酚对大鼠发育期大脑的神经毒性,部分机制可能是上调海马神经细胞抗凋亡蛋白Bcl-2和下调促凋亡蛋白Bax、cleaved-caspase-3水平。Objective To explore the effects of hypoxic preconditioning on neurotoxicity induced by propofol in neonatal rats.Methods Seventy⁃five 7⁃day⁃old SPF Sprague⁃Dawley rats,weighing 10‒15 g,were divided into five groups(n=15 in each group)using the random number table:a normal saline group(N group),a lipid emulsion group(F group),a propofol group(P group),a hypoxic preconditioning group(H group),and a hypoxic preconditioning+propofol group(H+P group).In the NS group,100μl normal saline was injected intraperitoneally;in the F group,100μl fat emulsion was injected intraperitoneally;in the P group,100 mg/kg pro⁃pofol was injected intraperitoneally;in the H group,rats were placed in 8%oxygen concentration for 10 min,air for 10 min,circulated for 5 times,and recovered in the air for 2 h;rats in the H+P group,received hypoxia pretreatment first(the same as H group)and then propofol was administered intraperitoneally(the same as P group).At the time of recovery from anesthesia,the rats were sacrificed to collect the hippocampus for the preparation of the specimen.The ultrastructure of hippocampal neurons was observed under a transmis⁃sion electron microscope.Paraffin sections were adopted for terminal deoxynucleotidyl transferase⁃mediated dUTP⁃biotin nick end la⁃beling(TUNEL)staining,and the number of positive cells was detected.The expression of cleaved⁃caspase⁃3,B⁃cell lymphoma⁃2(Bcl⁃2)and B⁃cell lymphoma⁃2⁃associated X protein(Bax)were determined by Western blot.Results Compared with the N group,the P group presented with shrunk hippocampal neurons,organelle disintegration and disappearance of the nucleoli,with an increased number of TUNEL positive cells(P<0.05),as well as increased expression of cleaved⁃caspase⁃3 and Bax protein(P<0.05)and de⁃creased expression of Bcl⁃2 protein(P<0.05).Compared with the P group,H+P group showed intact cellular membrane of hippocampal neurons,clear organelles and nuclei,and slightly reduced nuclear chromatin,with a decreased number of TUNEL p

关 键 词：新生大鼠 脑 神经毒性 缺氧预处理 异丙酚 

分 类 号：R72[医药卫生—儿科]