Th17及其细胞因子IL-17参与脑缺血损伤机制的研究

目的 检测Th17细胞在缺血脑组织中的表达及其细胞因子IL-17在厌氧条件下对神经元损伤机制的研究.方法 线栓法建立小鼠持续性大脑中动脉栓塞模型(pMCAO),免疫荧光技术检测pMCAO小鼠缺血侧脑组织中Th17细胞的表达；体外原代培养新生小鼠海马神经元,建立体外氧糖剥夺模型(OGD)模拟体内脑缺血缺氧微环境,观察不同厌氧时间对神经元的损伤作用,同时进行不同浓度的IL-17干预以及IL-17受体(IL-17R)封闭实验,探讨IL-17在厌氧条件下对神经元损伤过程中的作用.结果 成功建立小鼠右侧大脑中动脉栓塞模型,HE染色可见患鼠缺血侧脑组织空洞坏死样改变并有大量淋巴细胞浸润,TTC染色呈现白色缺血灶.免疫荧光染色可见缺血灶周边大量Th17细胞(CD4+IL-17+).体外实验中,外源性IL-17的干预加剧了OGD条件对海马神经元的损伤作用,并且这种作用能够被IL-17 R封闭剂所抑制.结论 小鼠缺血脑组织中存在Th17细胞的浸润,Th17细胞分泌的IL-17与厌氧条件下神经元的损伤有关.

Mechanism of Th17 and IL-17 in Cerebral Ischemic Injury

Objective To investigate the expression of Thl7 in isehemic brain tissue and the mechanism of IL-17 in potentiating neuronal injury induced by oxygen-glucose deprivation. Methods Set up the permanent middle cerebral artery occlusion （pMCAO）model by an intraluminal nylon introduction. Investigate the Thl7 cells in the ischemic brain tissue by immunofluorescence detection, oxygen-glucose deprivation （OGD） was induced in primary hippocampus cell cultures to modify isehemia microenvironment. Different dose of recombined IL-17 administration with or without IL-17R inhibitor was carried out to explore the mechanism of IL- 17 in potentiating neuronal injury under OGD. Results Animal model of pMCAO was successfully set up with large cerebral infarctions after Trc stain. Infiltration of Thl7 cells was observed according to the double-immu- nofluoreseence for CD4 and IL-17. Exogenous IL-17 facilitated OGD-mediated neuronal cell death which could be blocked using IL-17R inhibitor. Conclusion There was Th17 cells infiltration in the isehemie brain of pMCAO mice. IL-17 secreted from Th17 cells was relevant to OGD-indueed neuronal injury.