大鼠局灶性脑缺血再灌注后免疫蛋白酶体的变化  

Expression of Immunoproteasomes in the Pathogenesis of Focal Cerebral Ischemia in Rats

作　　者：陈兴泳[1] 汪银洲[1] 汪效松[1] 雷惠新[1] 唐荣华[2] 张旭[1] 

机构地区：[1]福建医科大学省立临床医学院福建省立医院神经内科,福州350001 [2]华中科技大学同济医学院附属同济医院神经内科,武汉430030

出　　处：《中国组织化学与细胞化学杂志》2015年第4期311-317,共7页Chinese Journal of Histochemistry and Cytochemistry

基　　金：福建省自然科学基金面上项目资助(2013J01275;2014J01401);福建省卫生厅中医药研究课题资助(wzzy201304)

摘　　要：目的观察大鼠局灶性脑缺血再灌注后脑组织免疫蛋白酶体LMP2和LMP7表达及其意义。方法线栓法制作SD大鼠大脑中动脉阻塞再灌注(middle cerebral artery occlusion,MCAO)模型,脑缺血1h再灌注72h。免疫荧光染色观察脑组织LMP2、LMP7、NF-κB、IL-1β、TNF-α表达和细胞分布。Western blot分析LMP2、LMP7、磷酸化NF-κB p65、IL-1β、TNF-α蛋白水平变化。结果 1局灶性脑缺血再灌注后上调LMP2和LPM7表达,尤其在梗死灶周边的皮层和纹状体区,与假手术组比较有显著性差异(P<0.001)。2免疫荧光双标显示星形胶质细胞是LMP2主要来源细胞,而OX42阳性的小胶质细胞/巨噬细胞是LMP7主要来源细胞;而且,NF-κB、IL-1β、TNF-α与LMP2、LMP7具有一定程度的细胞共定位。3Western blot结果表明,脑缺血再灌注后NF-κB p65、IL-1β、TNF-α蛋白水平表达趋势与LMP2、LMP7变化相类似。结论局灶性脑缺血再灌注后免疫蛋白酶体LMP2和LMP7主要来源于免疫炎症相关的细胞,推测LMP2和LMP7可能参与调节缺血性脑卒中后脑神经炎症反应。Objective To investigate the spatial expression and important role of immunoproteasomes LMP2 and LMP7in rat brains of the transient middle cerebral artery occlusion(MCAO)model.Methods MCAO was induced using the intraluminal suture occlusion technique in SD rats.Reperfusion of cerebral blood flow was allowed by gently removing the monofilament after 1-hour ischemia,followed by 72 hreperfusion.Immunofluorescent labeling and western blot were used to detect the expression of LMP2,LMP7,NF-κB,IL-1βand TNF-αin the brain,respectively.Results1 The expression of LMP2 and LMP7increased significantly in the ipsilateral ischemic hemisphere than that in the contralateral part at different time points,predominately in the cortex and striatum surrounding the infarct core(P<0.001,compared with sham-operated group).2Double immunostaining showed that LMP2 was mainly produced from astrocytes,whereas most of LMP7 was mainly produced by OX42-positive microglia/macrophage cells.Furthermore,NF-κB,IL-1βand TNF-αco-localized with LMP2 and LMP7,respectively.3Western blot confirmed that similarly to that of LMP2 or LMP7,the level of NF-κB p65,IL-1βand TNF-αproteins was upregulated after cerebral ischemia/reperfusion.Conclusion Both LMP2 and LMP7are mainly produced from inflammation cells after cerebral ischemia.Upregulation of LMP2 and LMP7potentially modulates post-ischemia neuroinflammation.

关 键 词：免疫蛋白酶体 神经炎症 脑缺血 

分 类 号：R743.3[医药卫生—神经病学与精神病学]