受体介导的1-磷酸鞘氨醇信号通路在局灶性脑缺血损伤中作用  

The function of receptor mediated sphingosine 1-phosphate signaling pathway in focal cerebral ischemia injury

作　　者：李洁琦 李萌 闵连秋 LI Jieqi;LI Meng;MIN Lianqiu(Department of Neurology,The First Affiliated Hospital of Jinzhou Medical University,Jinzhou 121000,China)

机构地区：[1]锦州医科大学附属第一医院神经内科,辽宁锦州121000 [2]抚顺市中心医院,辽宁抚顺113000

出　　处：《中风与神经疾病杂志》2019年第11期998-1003,共6页Journal of Apoplexy and Nervous Diseases

基　　金：辽宁省自然科学基金指导计划项目(No.20170540379)

摘　　要：目的探讨1-磷酸鞘氨醇(sphingosine 1-phosphate,S1P)受体亚型S1P3在局灶性脑缺血再灌注过程中的作用及其机制,为其作为药物靶点应用到脑缺血再灌注损伤的治疗提供理论依据。方法大脑中动脉闭塞法制备局灶性脑缺血再灌注的小鼠模型。用特异性S1P3拮抗剂CAY10444干预来评测S1P3在脑缺血再灌注中的作用。通过计算药物干预前后脑梗死体积、神经组织学评分和神经退行性变指标评估各组脑损伤程度。采用HE染色以及通过Iba1和Brd U/Iba1免疫组织化学染色方法观察和分析小胶质细胞的活化、形态转化和增殖情况。结果CAY10444抑制S1P3后显著降低MCAO诱导的脑梗死体积、神经功能缺损以及神经退行性变评分和程度。MCAO术后当S1P3活性受到抑制时,Iba1免疫阳性细胞的数量显著减少,脑缺血的核心区和周边区域活化的小胶质细胞数量均明显减少,同时,缺血核心区的小胶质细胞由阿米巴样形状恢复为分枝状形态。在MCAO后MCAO组脑缺血区小胶质细胞增殖明显,表现为Brd U/Iba1双免疫阳性细胞增多。与MCAO组相比,抑制S1P3后Brd U/Iba1双免疫阳性细胞的数量减少。结论S1P3是缺血再灌注脑损伤的致病介质,其机制可能是其参与调节了小胶质细胞活化和炎症发生,这为把S1P3作为脑缺血再灌注损伤的治疗靶点提供了依据。Objective To discuss the effect and its mechanismof receptor mediated sphingosine 1-phosphate(S1 P)signaling pathway in focal cerebral ischemia reperfusion,for the application to provide theoretical basis for treatment as drug targets.Methods Cerebral ischemia reperfusion in mice was reported by cerebral middle artery occlusion.The specific S1 P3 antagonist CAY10444 was used to evaluate the role of S1 P3 in cerebral ischemia reperfusion.Cerebral infarction volume,neurohistological score and neurodegenerative index were calculated before and after drug intervention to evaluate the degree of brain injury.The activation,morphological transformation and proliferation of microglia were observed and analyzed by HE staining and immunohistochemical staining with Iba1 and Brd U/Iba1.Results Inhibition of S1 P3 with CAY10444 immediately after reperfusion can significantly reduce MCAO induced cerebral infarction volume,neurological functional defects and neurodegeneration score.When S1 P3 activity was inhibited,the number of Iba1 immuno-positive cells was significantly reduced at the two time points above,and the number of activated microglias in the core area and peripheral area of cerebral ischemia was significantly reduced.After MCAO,the microglia in the ischemic core area recovered from amoeba-like shape to branched shape after S1 P3 inhibition.After MCAO,it was found that microglia cells in the cerebral ischemia area of the MCAO group had obvious proliferation,which was manifested as the increase of Brd U/Iba1 double-immune-positive cells.After MCAO,Compared with MCAO group,the number of Brd U/Iba1 double immuno-positive cells decreased significantly after S1 P3 inhibition.Conclusions S1 P3 is the pathogenic agent of cerebral ischemia/reperfusion injury,and it may be involved in the regulation of microglial cell activation and inflammation,which provides a basis for further using S1 P3 as a therapeutic target for cerebral ischemia/reperfusion injury in the future.

关 键 词：1-磷酸鞘氨醇 大脑中动脉闭塞 信号通路 脑缺血再灌注损伤 

分 类 号：R743[医药卫生—神经病学与精神病学]