CysLT_(2)受体拮抗剂HAMI3379对大鼠脑缺血损伤的保护作用及其机制  

Protective effect of CysLT_(2) receptor antagonist HAMI3379 on cerebral ischemia injury of rats and its mechanism

作　　者：李明星 应苗法[1] 顾胜龙 赵蕊[1] Li Ming-Xing;Ying Miao-Fa;Gu Sheng-Long;Zhao Rui(Department of Pharmacy,Sir Run Run Shaw Hospital,School of Medicine,Zhejiang University,Hangzhou,Zhejiang 310016,China)

机构地区：[1]浙江大学医学院附属邵逸夫医院药剂科,浙江杭州310016

出　　处：《解放军医学杂志》2023年第12期1395-1402,共8页Medical Journal of Chinese People's Liberation Army

基　　金：浙江省自然科学基金(LYY20H310002,LYY21H310002,LYY19H310010);浙江省中医药科技计划项目(2023ZF124)。

摘　　要：目的探究半胱氨酰白三烯2(CysLT_(2))受体拮抗剂HAMI3379对大鼠脑缺血损伤的保护作用及其可能机制。方法将30只雄性SD大鼠随机分为假手术组、模型组与HAMI3379组,每组10只。模型组大鼠采用大脑中动脉阻断术(MCAO)构建脑缺血损伤模型,HAMI3379组在MCAO术前和术后30min腹腔注射HAMI3379(0.2mg/kg)。对脑缺血损伤后的大鼠行神经症状评分,TTC染色法观察大鼠脑梗死体积,免疫荧光染色法检测小胶质细胞活化标志物Iba1,Real-time PCR检测小胶质细胞M1/M2极化表型分子mRNA水平的变化,NeuN染色法观察神经元数量,Fluoro-JadeB染色法观察神经元变性情况,Western-blotting检测脑组织中CysLT_(2)蛋白和核因子κB相关蛋白Cα(PKCα)、NF-κB抑制蛋白(IκBα)、p65和p50蛋白的表达变化。结果与假手术组比较,模型组大鼠神经症状评分、脑梗死体积均明显增高(P<0.05);与模型组比较,HAMI3379组大鼠的神经症状评分、脑梗死体积明显降低或缩小(P<0.05)。免疫荧光染色法检测结果显示,脑缺血损伤后大鼠脑组织中小胶质细胞活化标志物Iba1表达增加(P<0.05)。与假手术组比较,模型组大鼠缺血中心区脑组织中M1极化分子CD86、IL-1β、TNF-α和M2极化分子CD206、TGF-β、IL-10的mRNA表达均明显增高(P<0.05);与模型组比较,HAMI3379组M1极化分子表达明显下调(P<0.05),M2极化分子表达明显上调(P<0.05)。与假手术组比较,模型组大鼠脑组织中PKCα、IκBα、p65、p50蛋白表达明显上调(P<0.05);与模型组比较,HAMI3379组PKCα、IκBα、p65、p50蛋白表达明显下调(P<0.05)。NeuN染色结果显示,与假手术组比较,模型组大鼠脑组织中神经元数量减少(P<0.05),变性神经元数量增多(P<0.05);与模型组比较,HAMI3379组神经元数量增多(P<0.05),变性神经元数量减少(P<0.05)。结论CysLT_(2)受体拮抗剂HAMI3379可能通过调控PKCα/IκBα/NF-κB信号通路,抑制小胶质细胞的M1型极化激活,促进其向MObjective To explore the protective effects and mechanisms of cysteinyl leukotriene 2(CysLT_(2))receptor antagonist HAMI3379 on cerebral ischemia injury in rats.Methods 30 male SD rats were randomly divided into sham operation group,model group,and HAMI3379 group,with 10 rats in each group.The rats of model group were subjected to middle cerebral artery occlusion(MCAO)to construct the cerebral ischemia injury model,while HAMI3379 group received intraperitoneal injection of HAMI3379(0.2 mg/kg)before and after MCAO 30 min.The rats after cerebral ischemia injury were scored for neurological symptoms.The infarction volume of rats was observed by TTC staining,the activation marker Iba1 of microglia was detected by immunofluorescence staining,the mRNA level of M1/M2 polarized phenotype molecules of microglia was detected by Real-time PCR,the number of neurons was observed by NeuN staining,and neuronal degeneration was observed by Fluoro-Jade B staining.Western blotting assay was used to detect the expression of CysLT_(2) protein and nuclear factorsκB-related protein Cα(PKCα),IκBα,p65 and p50 proteins in brain tissue.Results Compared with sham operation group,the neurological symptom score and cerebral infarction volume of model group were significantly increased(P<0.05).Compared with model group,the neurological symptom score and cerebral infarction volume of HAMI3379 group were significantly decreased(P<0.05).The results of immunofluorescence staining showed that the expression of microglia activation marker Iba1 was increased in brain tissue of rats after cerebral ischemia injury(P<0.05).Compared with sham operation group,mRNA expression of M1 polarized molecules(CD86,IL-1β,TNF-α)and M2 polarized molecules(CD206,TGF-β,IL-10)were significantly increased in the ischemic central brain tissue of model group(P<0.05).Compared with model group,the expression of M1 polarized molecules in HAMI3379 group was significantly downregulated(P<0.05),while the expression of M2 polarized molecules was significantly upregulated

关 键 词：脑缺血损伤 CysLT_(2)受体 小胶质细胞激活 M1/M2极化 神经元损伤 

分 类 号：R743.3[医药卫生—神经病学与精神病学]