Col003对大鼠脑缺血再灌注损伤后血脑屏障的保护作用  

Protective effect of Col003 on the blood-brain barrier after cerebral ischemia-reperfusion injury in rats

作　　者：谢育良 周克剑 邓梦雨 谢晓云 梁婷婷 李浩英 刘竞丽[1] XIE Yuliang;ZHOU Kejian;DENG Mengyu;XIE Xiaoyun;LIANG Tingting;LI Haoying;LIU Jingli(Department of Neurology,the First Affiliated Hospital of Guangxi Medical University,Nanning 530021,China)

机构地区：[1]广西医科大学第一附属医院神经内科,南宁530021

出　　处：《广西医科大学学报》2025年第2期202-209,共8页Journal of Guangxi Medical University

基　　金：国家自然科学基金资助项目(No.82160236);广西自然科学基金资助项目(No.2023GXNSFDA026017)。

摘　　要：目的:探讨热休克蛋白47(HSP47)抑制剂Col003对大鼠脑缺血再灌注损伤后血脑屏障(BBB)的保护作用及其可能机制。方法:将大鼠随机分成假手术组、模型组、丁苯酞组、Col003组,采用线栓法构建大鼠大脑中动脉栓塞(MCAO)模型模拟脑缺血再灌注损伤,拔除线栓后丁苯酞组大鼠经腹腔注射丁苯酞(5 mg/kg),Col003组大鼠经尾静脉注射Col003(50μmol/L),两种药物的给药频次和持续时间均为每天1次,连续3 d,假手术组仅分离颈总动脉。采用蛋白免疫印迹法(western blotting)检测HSP47在大鼠脑缺血再灌注损伤后脑组织中的表达水平。通过TTC染色、神经运动功能评分、伊文思蓝渗透性实验评估大鼠脑功能障碍及BBB的损伤程度。通过western blotting检测大鼠脑组织中紧密连接蛋白(ZO-1、Occludin、Claudin5)以及间充质标志蛋白(a-SMA、Collagen I)的表达来观察Col003对大鼠脑缺血再灌注诱导的内皮—间质转化(EndMT)的影响,以进一步探讨Col003影响大鼠脑缺血再灌注后BBB损伤的可能机制。结果:在大鼠脑缺血再灌注后24 h、72 h,HSP47在脑组织中的表达水平均显著高于假手术组(P<0.0001)。与模型组比较,丁苯酞组和Col003组MCAO大鼠的神经功能评分(P<0.05)和运动功能评分(P<0.01)均得到显著改善,脑梗死面积均显著减小(P<0.0001),伊文思蓝渗透性均显著降低(P<0.0001)。与模型组比较,Col003组大鼠脑组织中紧密连接蛋白的表达显著增高(P<0.01),间充质标志蛋白的表达显著降低(均P<0.01)。结论:Col003可改善脑缺血再灌注后BBB的损伤,具有保护BBB的作用,其对BBB的保护作用可能是通过抑制EndMT实现。Objective:To investigate the protective effect of Col003,an inhibitor of heat shock protein 47(HSP47)on the blood-brain barrier(BBB)after cerebral ischemia-reperfusion injury in rats and its possible mechanism.Methods:The rats were randomly divided into sham group,model group,butylphthalide group,and Col003 group.The middle cerebral artery occlusion(MCAO)model in rats was constructed using the suture occlusion method to simulate cerebral ischemia-reperfusion injury.After removing the suture,the rats in the butylphthalide group were intraperitoneally injected with butylphthalide(5 mg/kg).The rats in the Col003 group were injected with Col003(50μmol/L)through the tail vein,and the frequency and duration of administration of both drugs were once a day for 3 consecutive days.In the sham group,only the common carotid artery was isolated.Western blotting was used to detect the expression level of HSP47 in the brain tissues of rats after cerebral ischemia-reperfusion injury.The degree of brain dysfunction and BBB damage was assessed using TTC staining,neurological motor function scores,and Evans blue permeability assays.The effects of Col003 on endothelialmesenchymal transformation(EndMT)induced by cerebral ischemia-reperfusion in rats were evaluated by detecting the expression of tight junction proteins(ZO-1,Occludin,Claudin5)and mesenchymal marker proteins(α-SMA,Collagen I)in rat brain tissues by using western blotting,so as to further explore the possible mechanism of Col003 affecting BBB injury after cerebral ischemia-reperfusion in rats.Results:At 24 h and 72 h after cerebral ischemia-reperfusion,the expression level of HSP47 in the brain tissue of rats was significantly higher than that in the sham group(P<0.0001).Compared with the model group,the neurological function scores(P<0.05)and motor function scores(P<0.01)of MCAO rats in butylphthalide group and Col003 group were significantly improved,the cerebral infarction area was significantly reduced(P<0.0001),and the Evans blue permeability was significantly dec

关 键 词：热休克蛋白47 Col003 缺血性卒中 缺血再灌注损伤 血脑屏障 内皮—间质转化 

分 类 号：R743.3[医药卫生—神经病学与精神病学]