4-PBA通过抑制内质网应激降低七氟醚诱导的HT22细胞凋亡  

4-PBA reduces the HT22 cell apoptosis after sevoflurane exposure by inhibiting stress endoplasmic reticulumstress

作　　者：祝刚[1] 陶蕾[2] 杨绍松[1] 王建才[1] 高国栋[1] 杨倩[1] 

机构地区：[1]第四军医大学唐都医院神经外科,西安710038 [2]第四军医大学唐都医院麻醉科,西安710038

出　　处：《神经解剖学杂志》2014年第6期634-638,共5页Chinese Journal of Neuroanatomy

基　　金：国家重点基础研究发展计划(973计划)(2011CB51000);国家自然科学基金面上项目(31371400);唐都医院科技创新发展基金重大国际合作项目(2013GJHZ001)

摘　　要：目的:探讨七氟醚暴露后的神经细胞损害以及通过4-苯基丁酸(4-phenyl butyric acid,4-PBA)抑制内质网应激对这种损害的保护作用。方法:以小鼠海马神经元来源的HT22细胞系为研究对象,将其分为对照组、七氟醚处理组、七氟醚+4-PBA预处理组。处理组在吸入性麻醉诱导箱中给予5%CO2和2%七氟醚麻醉处理5h,对照组只通入5%CO2,4-PBA预处理组则在麻醉前30 min给予4-PBA腹腔注射。检测内质网应激蛋白指标Bi P、p-IRE1以及活化的caspase12,并利用流式细胞法检测HT22细胞的凋亡情况。结果:(1)七氟醚诱导HT22细胞发生内质网应激,七氟醚处理后Bi P蛋白和IRE1的磷酸化水平明显升高,而4-PBA预处理组上述指标显著下降(P<0.01)。(2)七氟醚引起的HT22细胞凋亡能够被4-PBA抑制,流式细胞技术显示处理组细胞凋亡率明显大于4-PBA预处理组(P<0.01)。(3)内质网应激相关的caspase12激活介导了七氟醚引起的HT22细胞凋亡。统计学分析显示,处理组cleaved caspase12表达显著升高(P<0.01)。结论:吸入性麻醉剂七氟醚具有一定的神经毒性,内质网应激是其神经损害的重要机制之一。七氟醚能够通过诱导内质网应激而引起HT22细胞凋亡,而麻醉前给予4-PBA预处理可明显降低内质网应激反应,减少细胞凋亡,从而对七氟醚的神经毒性起到抵抗作用。Objective：To investigate the neuronal injury after sevoflurane exposure and the protection effect of 4-phenyl butyric acid（ 4-PBA） by inhibiting endoplasmic reticulum（ ER） stress.Methods：HT22 cells which were originated from mice hippocampal neuron were used in our present study and were divided into three groups,Control,Sevo and Sevo＋ 4-PBA.The Sevo group was exposed to 2% sevoflurane and 5% CO2 in an anesthesia chamber.Control group was exposed to 5% CO2 only and Sevo ＋ 4-PBA group was treated with 4-PBA 30 min before exposed to 2% sevoflurane and 5%CO2.Bi P and p-IRE1 which are the ER stress sensors and cleaved caspase12 were detected.Cell apoptosis in each group was detected by flow cytometry.Results：（ 1） Sevoflurane induced ER stress in HT22 cells,the levers of Bi P and pIRE1 significantly increased in Sevo group after sevoflurane exposure,and which were reduced in the Sevo ＋ 4-PBA group（ P〈0.01）.（ 2） 4-PBA inhibited sevoflurane caused cell apoptosis.Flow cytometry showed the greater apoptosis rate in Sevo group than Sevo ＋ 4-PBA groups（ P〈0.01）.（ 3） Cell apoptosis induced by sevoflurane was partly mediated by ERstress associated caspase12 activation.Statistical analysis shows the lever of cleaved caspase12 was elevated obviously in Sevo group（ P〈0.01）.Conclusion：Our research indicates that ER stress is a very important mechanism by which sevoflurane injures the neuronal system.Sevoflurane induced HT22 cell apoptosis by activating ER stress.Importantly,4-PBA protects HT22 cells from sevoflurane induced apoptosis by inhibiting ER stress.

关 键 词：七氟醚 内质网应激 4-苯基丁酸 凋亡 HT22细胞 

分 类 号：R614[医药卫生—麻醉学]