NADPH氧化酶2在七氟烷所致新生大鼠海马氧化应激及神经元凋亡中的作用机制  

The role of NADPH oxidase 2 in Sevoflurane-induced oxidative stress and apoptosis in the hippocampus of neonatal rats

作　　者：陈晓影[1] 蒋俊丹[1] 陈晓辉[1] Chen Xiaoying;Jiang Jundan;Chen Xiaohui(Department of Anesthesiology,Fujian Provincial Hospital,Fuzhou 350001,China)

机构地区：[1]福建省立医院麻醉科,福州350001

出　　处：《创伤与急诊电子杂志》2023年第2期53-61,共9页Journal of Trauma and Emergency(Electronic Version)

基　　金：国家自然科学基金青年科学基金项目(82001166)。

摘　　要：目的探究吸入性麻醉药七氟烷是否通过激活新生大鼠海马NADPH氧化酶2介导氧化应激损伤引起神经元凋亡及远期认知功能损害。方法新生7d的清洁级SD雄性大鼠116只,采用随机数字表法分为空气对照组(Con组,n=29)、夹竹桃麻素对照组(Apo组,n=29)、七氟烷实验组(Sevo组,n=29)、七氟烷+夹竹桃麻素实验组(Sevo+Apo组,n=29)。Con组和Sevo组新生大鼠分别暴露于空气或2.3%七氟烷中6h,Apo组和Sevo+Apo组于空气或七氟烷暴露前腹腔注射给予新生大鼠夹竹桃麻素50mg/kg。于七氟烷暴露结束后6h检测4组新生大鼠海马组织NADPH氧化酶2活性和蛋白表达水平,评估氧化应激产物丙二醛(malondialdehyde,MDA)含量和凋亡相关蛋白cleaved caspase-3表达水平,运用尼氏染色法观察海马CA1、CA3区神经元的形态学变化并计数,并于七氟烷暴露结束后21d采用八臂迷宫实验检测大鼠空间工作记忆和参考记忆用于评估学习记忆能力变化。结果与Con组比较,Sevo组NADPH氧化酶2活性及蛋白表达水平均升高(P<0.05),氧化应激产物MDA含量及凋亡相关蛋白cleaved caspase-3表达水平同样升高(P<0.05);而与Sevo组相比,Sevo+Apo组NADPH氧化酶2活性及蛋白表达量、MDA含量及cleaved caspase-3表达水平均降低(P<0.05)。与Con组比较,Sevo组海马锥体神经元排列明显稀疏紊乱,部分胞质溶解,细胞核固缩、碎裂伴有CA1区和CA3区锥体神经元数目减少(P<0.05),Sevo+Apo组也可见细胞水肿、核固缩等病理形态,但其程度较Sevo组明显减轻,且椎体神经计数仍多于Sevo组(P<0.05)。八臂迷宫实验提示第1~5天Sevo组工作记忆错误次数高于Con组(P<0.05),但Sevo+Apo组出现空间工作记忆错误次数少于Sevo组,差异有统计学意义(P<0.05);而在参考记忆错误方面,4组大鼠表现差异无统计学意义(P>0.05)。结论七氟烷所致发育脑海马神经元凋亡及认知功能损害与其激活NADPH氧化酶2介导氧化应激损伤密切相Objective To explore whether inhaled anesthetic Sevoflurane causes neuronal apoptosis and long-term cognitive impairment in neonatal rats by triggering oxidative stress through the activation of the hippocampal NADPH oxidase 2.Method A total of 116 male SD rats(7 days old,clean grade)were randomly assigned to four groups:control(Con)group,apocynin-treated(Apo)group,Sevoflurane-treated(Sevo)group and Sevoflurane+apocynin(Sevo+Apo)group.The neonatal rats in the Con and Sevo groups were exposed to air or 2.3%Sevoflurane for 6 hours,while the rats in the Apo and Sevo+Apo groups were injected with 50mg/kg of apocynin into the peritoneal cavity before being exposed to air or Sevoflurane.At 6 hours following sevoflurane exposure,the activity and protein expression levels of NADPH oxidase 2 in the hippocampus of the four groups of rats were measured,the levels of oxidative stress products MDA and the expression levels of apoptotic protein cleave caspase-3,and the morphological changes and cell counting of the hippocampal neurons in CA1 and CA3 regions were examined.Twenty-one days after sevoflurane exposure,the spatial working memory and reference memory were evaluated by using the eight-arm maze test.Result Compared with the Con group,the Sevo group showed a significant increase in NADPH oxidase 2 activity and protein expression level(P<0.05),as well as an increase in MDA level and cleaved caspase-3 expression level(P<0.05).Compared with the Sevo group,the Sevo+Apo group showed a decrease in NADPH oxidase 2 activity and protein expression level,as well as a decrease in MDA level and cleaved caspase-3 expression level(P<0.05).The Sevo group exhibited a significant decrease in the number of hippocampal pyramidal neurons in CA1 and CA3 regions,as well as a disordered arrangement of neurons,partial cytoplasmic dissolution,and nucleus shrinkage and fragmentation(P<0.05);whereas the Sevo+Apo group showed a milder pathological change,and the number of pyramidal neurons was still higher than that in the Sevo group(P<0.05).In th

关 键 词：七氟烷 新生大鼠 海马 NADPH氧化酶2 氧化应激 凋亡 

分 类 号：R614[医药卫生—麻醉学]