急性颈动脉血栓性闭塞形成后巨噬细胞浸润行为的动态演变研究  

A study of the dynamic evolution of macrophage infiltration behavior after acute carotid artery thrombosis

作　　者：李世凯 梁佳 何艳艳 丁乾昊 李晨卿 刘阳 冯英璞 赵文利 黄宇梦 索莉娜 李天晓 贺迎坤 Li Shikai;Liang Jia;He Yanyan;Ding Qianhao;Li Chenqing;Liu Yang;Feng Yingpu;Zhao Wenli;Huang Yumeng;Suo Lina;Li Tianxiao;He Yingkun(School of Medical Engineering,Xinxiang Medical University,Xinxiang,Henan 453003,China;不详)

机构地区：[1]新乡医学院医学工程学院,453003 [2]河南省脑血管病国际联合实验室、河南省脑血管介入创新工程技术研究中心 [3]河南省人民医院(郑州大学人民医院)脑血管病科

出　　处：《中国脑血管病杂志》2024年第7期433-443,共11页Chinese Journal of Cerebrovascular Diseases

基　　金：国家卫生健康委能力建设和继续教育中心2023年度立项课题(GWJJ2023100101);河南省重点研发专项(241111313200);河南省科技攻关计划项目(232102231042);河南省医学科技攻关计划省部共建青年项目(SBGJ202303001)。

摘　　要：目的探索急性颈动脉血栓性闭塞(ACTO)形成后,血栓动态演变过程中巨噬细胞的浸润行为变化。方法选用健康雄性新西兰大白兔15只,通过使用氯化铁处理后的外科缝合线对兔颈动脉造成损伤来建立ACTO模型。采用随机数字表法将所有兔按照实验终点时间分为5组;术后24 h组及术后1、4、8、12周组,每组3只。术后24 h,使用DSA检查ACTO模型建立情况。到达实验时间终点时对该组3只兔进行血栓栓塞段动脉取材并用石蜡包埋。采用苏木素-伊红(HE)染色和马休黄猩红蓝(MSB)对血栓样本进行染色,分析血栓形态和血栓组成(包括红细胞、纤维蛋白和胶原纤维)的变化,使用Orbit Imaging Analysis软件对血栓组成成分进行半定量分析。使用免疫组织化学对血栓中M0和M2型巨噬细胞的分布进行检测,依据检测结果总结ACTO发生后不同时期血栓的形态、组成及血栓中巨噬细胞浸润行为的动态演变规律。结果术后24 h DSA结果显示,所有的实验兔均成功建立了ACTO模型。(1)HE染色结果显示,在术后24 h至1周的时间段内,血栓呈持续增大趋势;术后4周时,血栓开始出现溶解现象;术后8周时,血栓内部观察到了新生血管的形成;而术后12周时,血栓开始呈现出纤维化的迹象。(2)MSB染色结果显示,在血栓形成的急性期(术后24 h内),红细胞为主要成分;在此之后,以纤维蛋白和胶原纤维为主要成分。(3)M0型巨噬细胞(细胞表面标志物为CD206)检测结果显示,术后24 h,血栓中M0型巨噬细胞并未均匀分布于整个血栓中,而主要聚集于血栓边缘;术后1周时,血栓内M0型巨噬细胞阳性面积百分比相较于术后24 h增加[血栓边缘:(41.7±27.0)%比(24.6±16.7)%,血栓核心:(35.7±19.6)%比(11.1±10.4)%,均P<0.001],且均匀分布于血栓内;术后4周时,血栓内M0型巨噬细胞的阳性面积百分比相较于术后1周减少[血栓边缘:(10.7±6.1)%比(41.7±27.0)%,血栓核心:(12.1±8.5)%比(3Objective To explore the changes in macrophage infiltration behavior during the dynamic evolution of thrombi following the formation of acute carotid artery thrombosis occlusion(ACTO).Methods 15 healthy male New Zealand rabbits were selected to establish an ACTO model by causing injury to the rabbit carotid artery using surgical sutures treated with ferric chloride.All rabbits were randomly divided into 5 groups according to the end-point time using the random number table method,namely 24-hour group,1 week group,4 week group,8 week group,and 12 week group postoperatively,with 3 rabbits in each group.At 24 hours post-operation,the ACTO model was examined by DSA.At 24 hours,1 week,4 weeks,8 weeks,and 12 weeks post-operation,samples were taken from the thrombotic arterial segment of the 3 rabbits in each group and embedded in paraffin.The thrombus samples were stained with hematoxylin-eosin(HE)and Martius scarlet blue(MSB)to analyze changes in thrombus morphology and composition(including red blood cells,fibrin and collagen fibers).Orbit Imaging Analysis software was used for semi-quantitative analysis of the thrombus composition components.Using immunohistochemistry to detect the distribution of M0 and M2 macrophages in thrombi,aimed to summarize the dynamic evolution of thrombus morphology,composition,and macrophage infiltration behavior at different stages following ACTO occurrence.Results The 24-hour DSA results indicated that all experimental rabbits successfully established the ACTO model.(1)HE staining showed a continuous increase in thrombus size from 24 hours to 1 week.By 4 weeks,signs of thrombus dissolution appeared,and at 8 weeks,neovascularization was observed within the thrombus.By 12 weeks,signs of fibrosis were evident in the thrombus.(2)MSB staining revealed that during the acute phase of thrombus formation(within 24 hours after surgery),red blood cells were the predominant component initially,but after this period,fibrin and collagen fibers became the main components.(3)The detection results of M0

关 键 词：急性颈动脉血栓 血栓栓塞 巨噬细胞 动物模型 

分 类 号：R743[医药卫生—神经病学与精神病学]