基于脑小血管病临床脑血流特点构建并评估不完全性全脑缺血再灌注大鼠模型  

Construction and evaluation of an incomplete global cerebral ischemia-reperfusion rat model based on hemodynamic characteristics of cerebral small vessel disease

作　　者：王珊珊[1,2] 徐昊 侯培媚[2,3] 李泽康 周丽娟 葛金文 WANG Shanshan;XU Hao;HOU Peimei;LI Zekang;ZHOU Lijuan;GE Jinwen(College of Integrated Chinese and Western Medicine,Hunan University of Chinese Medicine,Changsha 410208,China;Key Laboratory of Hunan Province for Integrated Chinese and Western Medicine on Prevention and Treatment of Cardio-Ce-rebral Diseases,Hunan University of Chinese Medicine,Changsha 410208,China;College of Acupuncture,Massage and Rehabilitation,Hunan University of Chinese Medicine,Changsha 410208,China;College of Pharmacy,Hunan Universi-ty of Chinese Medicine,Changsha 410208,China)

机构地区：[1]湖南中医药大学中西医结合学院,湖南长沙410208 [2]湖南中医药大学中西医结合防治心脑血管疾病湖南省重点实验室,湖南长沙410208 [3]湖南中医药大学针灸推拿与康复学院,湖南长沙410208 [4]湖南中医药大学药学院,湖南长沙410208

出　　处：《中国病理生理杂志》2023年第7期1330-1338,共9页Chinese Journal of Pathophysiology

基　　金：湖南省自然科学基金资助项目(No.2020JJ5424);湖南省教育厅青年基金资助项目(No.21B0386);湖南中医药大学中西医结合“双一流”学科开放基金资助项目(No.2020ZXYJH08)。

摘　　要：目的:基于脑小血管病(CSVD)的脑血流特点构建和评估不完全性全脑缺血再灌注大鼠模型,以期为CSVD等缺血性脑血管疾病的基础研究提供理想的大鼠实验模型。方法:将126只雄性SD大鼠随机分为假手术(sham)组、轻损伤模型组(minor组)和重损伤模型组(serious组),每组42只。采用双侧颈总动脉夹闭-开放循环操作的方法,构建大鼠不完全性全脑缺血再灌注模型,激光散斑血流成像系统评估脑血流实时变化;于造模后2、24和72 h,评估造模动物存活率和动物行为学(平衡木实验,BBT)评分;于造模2、24和72 h取材,采用HE染色观察大鼠不同脑区组织病理形态改变;并运用非靶向代谢组学分析模型大鼠血浆和脑皮质区差异代谢物,评估模型损伤程度。结果:(1)与sham组比较,minor组和serious组大鼠总存活率平均值分别为83.2%和73.7%(P<0.05);(2)残存脑血流量平均值分别为56.3%和40.9%;(3)神经功能检查显示,与sham组比较,minor组和serious组造模后2、24和72 h的BBT评分均显著升高(P<0.05);(4)HE染色镜下可见广泛脑皮质(M1区和RSA区最为明显)、腹内测下丘脑腹外侧部(VMHvl)和海马C2区神经细胞形态发生不同程度改变。(5)非靶向代谢组学结果显示,sham组和serious组大鼠血浆差异性代谢物总数45个,上调代谢物总数33个,下调代谢物总数12个;脑皮质RSA区差异性代谢物总数19个,上调代谢物总数6个,下调代谢物总数13个,提示造模导致了大鼠神经-内分泌功能的紊乱。结论:双侧颈总动脉夹闭-开放循环造模方法可导致大鼠脑皮质、VMHvl和海马C2区散在性广泛损伤,其损伤机制可能与代谢紊乱、氧化应激损伤等有关。该模型为研究CSVD反复缺血再灌注损伤提供了新的大鼠实验模型。AIM:To construct and evaluate an incomplete global cerebral ischemia-reperfusion rat model based on the hemodynamic characteristics of cerebral small vessel disease(CSVD),so as to provide an ideal rat model for investigating ischemic cerebrovascular diseases such as CSVD.METHODS:A total of 126 male SD rats were divided into sham,minor,and serious groups(n=42).The bilateral common carotid artery clamp-open circulation operation was used to construct the incomplete global cerebral ischemia-reperfusion rat model,and real-time changes of cerebral hemodynamics were assessed by a laser speckle blood flow imaging system.At 2,24 and 72 h after the model was constructed,the survival rate and animal behavior(beam balance test,BBT)score were evaluated,and the materials of the model animals were taken.HE staining was used to observe the histopathological changes in different brain regions of the rats,and non-targeted metabonomics was utilized to analyze the different metabolites in the rat plasma and cerebral cortex,thus evaluating the damage degree.RESULTS:The average survival rates of rats in minor and serious groups were 83.2%and 73.7%(P<0.05),and the mean residual cerebral blood flow rates were 56.3%and 40.9%,respectively.The BBT scores at 2,24 and 72 h after modeling in minor and serious groups were higher than those in sham group(P<0.05).The HE staining results showed that the neurons in the extensive cerebral cortex(especially in M1 and RSA),ventrolateral part of ventromedial hypothalamus(VMHvl)and hippocampal C2 region showed morphological changes at different degrees.The non-targeted metabonomics analysis revealed a total of 45 differential metabolites in the plasma of sham and serious groups,comprising 33 up-regulated and 12 down-regulated metabolites.In addition,the RSA exhibited 19 differential metabolites,including 6 up-regulated and 13 down-regulated metabolites.This finding suggests that the modeling disrupted neuroendocrine function in the rats.CONCLUSION:The bilateral common carotid artery occlusion and open

关 键 词：脑小血管病 缺血性脑卒中 缺血再灌注损伤 动物模型 

分 类 号：R363[医药卫生—病理学] R743[医药卫生—基础医学] R-33