安菲博肽调控小胶质细胞M1/M2极化抑制脑缺血再灌注损伤后炎症反应研究  

Inhibition Effect of Anfibatide on Inflammation after Cerebral Ischemia Reperfusion Injury through Regulating M1/M2 Microglia Polarization

作　　者：宫继鹏[1] 贾会玉 李睿[2] 马征[2] 李心伟[2] 司敏 戴向荣 罗胜勇[2] CONG Ji-peng;JIA Hui-yu;LI Rui;MA Zheng;LI Xin-wei;SI Min;DAI Xiang-rong;LUO Sheng-yong(The First Afiliated Hospital of Anhui Medical University,Hefei 230032,China;Anhui Provincial Institute of Medical Scince,Hefei 230061,China;Zhaoke Pharmaceutical(hefei)Company Limited,Hefei 230088,China)

机构地区：[1]安徽医科大学第一附属医院,合肥230032 [2]安徽省医学科学研究院,合肥230061 [3]兆科药业(合肥)有限公司,合肥230088

出　　处：《中国药学杂志》2023年第5期426-434,共9页Chinese Pharmaceutical Journal

基　　金：安徽省自然科学基金资助(2108085MH322)。

摘　　要：目的研究安菲博肽(anfibatide,ANF)对脑缺血再灌注损伤后小胶质细胞M1/M2极化及神经炎症的影响。方法取SD大鼠64只,随机分为正常组,模型组,Toll样受体4(TLR4)阻断剂TAK-242(2 mg·kg^(-1)),ANF(4μg·kg^(-1))组,每组16只。线栓法建立大鼠局灶性脑缺血再灌注损伤模型,缺血2 h后恢复再灌,连续尾静脉注射给药5 d。氯化三苯基四氮唑(TTC)染色测定脑梗死体积,神经功能缺损评分(mNSS)方法进行神经功能缺损评分;免疫荧光及Western blot测定脑组织中CD86/CD206蛋白表达。取BV-2细胞,建立糖氧剥夺/复糖复氧(OGD/R)模型,分对照组、OGD/R组、TAK-2421μmol·L^(-1)组和ANF 0.1、0.2、0.4μg·mL^(-1)组,缺糖缺氧6 h后,复糖复氧24 h并用相应药物进行干预。免疫荧光及实时定量PCR(RT-PCR)测定各组BV-2细胞CD86/CD206蛋白及肿瘤坏死因子-α(TNF-α)、白细胞介素-1β(IL-1β)、IL-10、转化生长因子-β(TGF-β)基因表达。结果与模型组比较,ANF可明显减少脑梗死体积,改善大鼠神经功能缺损,明显降低脑组织中CD86蛋白表达,明显升高CD206蛋白表达。与对照组比较,ANF 0.1、0.2、0.4μg·mL^(-1)可明显降低BV-2中CD86免疫荧光表达,明显降低CD86、IL-1β基因表达,明显升高CD206基因表达,0.2、0.4μg·mL^(-1)可明显升高CD206免疫荧光表达,明显降低TNF-α表达;升高IL-10、TGF-β基因表达水平。结论ANF可通过抑制小胶质细胞M1极化、促进M2极化,从而减轻脑缺血再灌注损伤后神经炎症反应。OBJECTIVE To study the effect of anfibatide(ANF)on microglia M1/M2 polarization and neuroinflammation af-ter cerebral ischemia/reperfusion injury.METHODS Sixty-four SD rats were randomly divided into four groups:sham,model,TAK-242(2 mg·kg^(-1),antagonist of tll-like receptor 4,TLR4),and ANF 4μg kg^(-1)group.The occlusion model of mid-dle cerebral artery(MCAO)was established by thread embolism method.Reperfusion was resumed after 2 h of ischemia,and the drug was administered continuously by tail vein for 5 d.The neurological deficit was evaluated by mNSS and the infarct volume was measured by TTC staining.The expressions of CD86,CD206 proteins in brain tissue were determined by immunofluores-cence and Western-blot method.BV-2 cells were used to establish OGD/R model and divided into control,OGD/R,TAK-2421μmol·L^(-1),ANF 0.1,0.2,0.4μg·mL^(-1) group.After 6 h of hypoxia,reoxygenation was performed for 24 h and corre-sponding drugs were used for intervention.The expressions of CD86,CD206 proteins were determined by immunofluorescence,and CD86,CD206,TNF-α,IL-1β,IL-10 and TCF-βgenes in BV-2 cells were detected by real-time quantitative PCR(RT-PCR).RESULTS Compared with the model group,ANF could significantly reduce the cerebral infarction volume,decrease the neurological deficit and the expression of CD86 protein in brain tissue,and significantly increase the expression of CD206 pro-tein.In vitro,compared with the control group,ANF 0.1,0.2,0.4μg·mL^(-1) significantly decreased the immunofluorescence expression of CD86 in BV-2,significantly reduced the expression of CD86,IL-1βmRNA,obviously increased the expression of CD206 mRNA,and ANF 0.2,0.4μg·mL^(-1) obviously increased the immunofluorescence expression of CD206.Furthermore,

关 键 词：安菲博肽 脑缺血再灌注 炎症 小胶质细胞 极化 

分 类 号：R965[医药卫生—药理学]