脑缺血损伤介导星形胶质细胞血小板激活因子受体变化的初探  

Exploration of Platelet Activating Factor Receptor in Astrocytes after Cerebral Ischemia

作　　者：范国航 韩思达 肖保国[2] 董强 FAN Guo-hang;HAN Si-da;XIAO Bao-guo;DONG Qiang(Department of Neurology,Huashan Hospital,Fudan University,Shanghai 200040,China;Institute of Neurology,Huashan Hospital,Fudan University,Shanghai 200040,China)

机构地区：[1]复旦大学附属华山医院神经内科,上海200040 [2]复旦大学神经病学研究所,上海200040

出　　处：《中国临床神经科学》2022年第4期361-368,共8页Chinese Journal of Clinical Neurosciences

基　　金：国家自然科学基金委面上项目(编号:81870915)。

摘　　要：目的探究脑缺血损伤后星形胶质细胞血小板激活因子(PAF)受体(PAFR)的变化及其诱导因素。方法利用体外培养的原代星形胶质细胞建立氧糖剥夺/复氧(OGD/R)细胞模型;制备小鼠大脑中动脉梗死(MCAO)模型。观察OGD/R和MCAO后不同时程星形胶质细胞PAFR表达水平的变化。分别采用体外H2O2与PAF处理原代星形胶质细胞,予小鼠侧脑室注射不同剂量PAF进行干预,探索缺血损伤后PAFR表达的影响因素和PAFR上调后发挥的生物学功能。用免疫荧光法检测PAFR和PAF的表达水平,Western blot法检测PAFR、凋亡相关蛋白表达水平。结果经OGD/R处理的星形胶质细胞PAFR表达水平显著上升;MCAO后3 d,模型小鼠星形胶质细胞PAFR表达水平明显增加,7 d后PAFR表达水平有所降低。经H2O2处理的星形胶质细胞PAFR较对照组表达水平明显增加。内源性PAF水平以MCAO后1 d最高,随着时间延长急剧降低,MCAO后3 d近乎基线水平。侧脑室注射PAF与体外PAF干预星形胶质细胞均可诱导PAFR表达。凋亡相关蛋白caspase-3和Bax表达随着PAFR水平的增加而上调,提示PAFR与细胞凋亡存在密切关系。结论缺血缺氧诱导星形胶质细胞PAFR表达,缺血损伤引起的氧化应激损伤和内源性PAF释放是重要原因。PAFR增加与缺血性脑卒中后细胞凋亡存在密切联系,精准调控PAFR可能成为缺血性脑卒中神经保护的有效策略。Aim To investigate the changes of astrocyte platelet activating factor receptor(PAFR)expression and its triggering factors under cerebral ischemia.Methods Primary astrocytes cultured in vitro were used to establish the oxygen glucose deprivation/reoxygenation(OGD/R)model,and mouse was utilized to prepare middle cerebral artery infarction(MCAO)model.The changes of PAFR expression in astrocytes after OGD/R and MCAO were observed by Western blot and immunocytochemistry staining.PAF stimulation and H2O2 treatment in vitro were used to understand the possible triggering factors of PAFR induction in OGD/R model.PAF injection through lateral ventricle provided direct evidence of PAF-activated PAFR expression.Results In OGD/R model,the expression of PAFR in astrocytes increased significantly.Three days after MCAO,the expression of PAFR in astrocytes increased and then decreased 7 days after MCAO,while the expression of PAF increased rapidly after one day of MCAO.H2O2 treatment in vitro directly induced the expression of PAFR in astrocytes.PAF stimulation upregulated PAFR in vitro as well.Intracerebroventricular PAF injection further confirmed that PAF could specifically induce the expression of PAFR receptor in astrocytes.PAFR was accompanied by the increased expression of apoptotic proteins caspase-3 and Bax,suggesting PAFR may relate to cell apoptosis.Conclusion The expression of PAFR in astrocytes was induced by OGD/R and MCAO,which should be related to PAF and oxidative stress.PAFR was involved in the pathological process of ischemic stroke,and may become a potential target for neuroprotective treatment and drug development in cerebral ischemic stroke.

关 键 词：缺血性脑卒中 星形胶质细胞 血小板激活因子受体 氧化应激 神经保护 

分 类 号：R743[医药卫生—神经病学与精神病学] Q189[医药卫生—临床医学]