二十二碳六烯酸对七氟醚诱导的小胶质细胞Toll样受体4/髓样分化因子88/核因子-kB信号通路激活及炎症介质释放的影响  

Effect of docosahexenoic acid on sevoflurane-induced activation of microglia Toll 4ike receptor 4/myeloid differentiation factor 88/nuclear factor-ĸB pathway and the release of inflammatory mediators

作　　者：赵敏 赵品 葛娜 张尚民 蒯建科 Zhao Min;Zhao Pin;Ge Na;Zhang Shangmin;Kuai Jianke(Department of Anesthesiology,Xi'an No.3 Hospital,Xi'an 710003,China;Department of Anesthesiology,Xi'an Angel Women’s Hospital,Xi'an 710021,China)

机构地区：[1]西安市第三医院麻醉科,71OOO3 [2]西安安琪儿妇产医院麻醉科,710021

出　　处：《国际麻醉学与复苏杂志》2019年第12期1134-1138,共5页International Journal of Anesthesiology and Resuscitation

基　　金：国家自然科学基金(81470414)。

摘　　要：目的观察二十二碳六烯酸(docosahexenoic acid,DHA)对七氟醚诱导的小胶质细胞Toll样受体4(Toll-like receptor 4,TLR4)/髓样分化因子88(myeloid differentiation factor 88,MyD88)/NF-ĸB信号通路激活及炎症介质释放的影响。方法N9小鼠小胶质细胞分为对照组(CON组)、七氟醚组(Sevo组)、七氟醚和DHA处理组(DHA+Sev组),药物处理各组细胞24 h后,采用噻唑蓝(methylthiazolyldiphenyl-tetrazolium bromide,MTT)法检测小胶质细胞存活率,采用Western blot法检测小胶质细胞TLR4、MyD88和.NF-ĸB抑制蛋白-α(inhibitor of NF-ĸB-α,IĸB-α)的表达量,采用ELISA法检测各组培养基中炎症介质TNF-α、IL-1β、IL-6和IL-10的含量。结果与CON组比较,Sevo组细胞存活率降低,TLR4和MyD88表达水平升高,IĸB-α表达水平降低,TNF-α、IL-1β和IL-6释放量升高(P<0.05),DHA+Sevo组细胞IL-1β、IL-10释放量升高(P<0.05);与Sevo组比较,DHA+Sevo组小胶质细胞存活率升高,TLR4和MyD88表达水平明显降低,IĸB-α表达水平升高,TNF-α、IL-1β和IL-6释放量降低,而IL-10释放量升高(P<0.05)。结论DHA抑制七氟醚所致细胞损伤以及TLR4/MyD88/NF-ĸB信号通路激活,并且减少炎症介质TNF-α、IL-1β和IL-6的释放,增加IL-10释放量。Objective To observe the effects of docosahexenoic acid(DHA)on sevoflurane-induced Toll-like receptor 4(TLR4)/myeloid differentiation factor 88(MyD88)/nuclear factor-ĸB(NF-ĸB)pathway activation and the release of inflammatory mediators.Metheds N9 cells were assigned to a CON group,a Sevo group and a DHA+Sevo group.After 24h treatment,cell survival was assessed by methylthiazolyldiphenyl-tetrazolium bromide(MTT)assay.Western blot was used to detect the levels of microglial TLR4,MyD88 and inhibitor of NF-ĸB-α(IĸB-α),while the contents of inflammatory mediator tumor necrosis factor-α(TNF-α),interleukin(IL)-1β,IL-6 and IL-10 in each group were detected by enzyme-linked immunosorbent assay(ELISA).Results Compared with the CON group,the survival rate of N9 cells in the Sevo group decreased significantly,while the level of TLR4 and MyD88 protein was increased significantly,the level of IĸB-αwas significantly decreased,the release of TNF-αand IL-1βand IL-6 was increased significantly(P<0.05).The amounts of IL-1βand IL-10 were increased significantly in the DHA+Sevo group(P<0.05).Compared with the Sevo group,the survival rate in the DHA+Sevo group increased significantly,the level of TLR4 and MyD88 protein decreased significantly,the level of IĸB-αwas increased significantly,TNF-α,IL-1βand IL-6 release was increased significantly,while IL-10 release was increased significantly(P<0.05).Conclusions DHA inhibits sevoflurane-induced cell damage and activation of TLR4/MyD88/NF-ĸB signaling pathway,and reduces the release of pro-inflammatory mediators TNF-α,IL-1βand IL-6,and increases anti-inflammatory mediator IL-10 release.

关 键 词：二十二碳六烯酸 七氟醚 小胶质细胞 Toll样受体4/髓样分化因子88/核因子-ĸB信号通路 炎症介质 

分 类 号：R614[医药卫生—麻醉学]