摘要
目的建立并评估能模拟外源性物理损伤的颈总动脉钳夹致颈总动脉钝性创伤模型。方法取64只健康雄性C57BL/6J小鼠随机分为假手术组和颈总动脉钳夹致颈总动脉钝性创伤建模后1、3、7 d组,每组16只。假手术小鼠予颈部切开、缝合,建模小鼠予左侧颈总动脉钳夹30 min。取颈总动脉标本,通过H-E染色观察血管组织结构改变,免疫组织化学染色(n=10)、蛋白质印迹法(n=6)检测血管功能相关蛋白的表达。结果H-E染色显示颈总动脉钳夹伤后血管内皮细胞出现不同程度损伤,血管基质的连续性下降。免疫组织化学染色显示颈总动脉钳夹伤后血管细胞黏附分子1(VCAM-1)、内皮型一氧化氮合酶(eNOS)、内皮素1表达上调,炎症细胞标志物髓过氧化物酶(MPO)在血管壁及血管周围表达增加。蛋白质印迹法检测结果显示,颈总动脉钳夹伤后血管功能相关标志物VCAM-1、eNOS、内皮素1,炎症相关蛋白环氧化酶2、基质金属蛋白酶2、基质金属蛋白酶9,以及凋亡相关蛋白B淋巴细胞瘤2相关X蛋白、活化caspase 1的表达均有不同程度上调,其中血管功能相关标志物VCAM-1与eNOS表达均在钳夹后3 d到达高峰,而内皮素1表达则在钳夹后1~7 d呈上升趋势。结论颈总动脉钳夹致颈总动脉钝性创伤小鼠模型建立成功。颈总动脉钳夹可导致血管组织损伤并发生炎症反应与细胞凋亡,这种损伤可能是创伤性血管狭窄及动脉瘤形成的重要因素。
Objective To establish and evaluate a blunt common carotid artery trauma model created by common carotid artery clipping,which can simulate exogenous physical injury.Methods Sixty-four healthy male C57BL/6J mice were randomly divided into sham group,and 1,3 and 7 d after clipping groups,with 16 mice in each group.The mice in the sham group were given cervical incision suture,and those in the modeling group were given left common carotid artery clipping occlusion for 30 min.Common carotid arteries were then harvested,the vascular structure was observed by hematoxylin-eosin(H-E)staining,and the expression of vascular function related proteins was detected by immunohistochemical staining(n=10)and Western blotting(n=6).Results H-E staining showed that the vascular endothelial cells were injured to different extents,and the integrity of vascular matrix was decreased after carotid artery clipping.Immunohistochemical staining showed that the expression levels of vascular cell adhesion molecule 1(VCAM-1),endothelial nitric oxide synthase(eNOS)and endothelin-1 were up-regulated after common carotid artery clipping,and the expression of inflammatory cell marker myeloperoxidase(MPO)was also elevated in vascular walls and perivascular tissues.The results of Western blotting showed that the expression levels of vascular function related markers(VCAM-1,eNOS and endothelin-1),inflammation-related proteins(cyclooxygenase 2[COX-2],matrix metalloproteinase[MMP]-2 and MMP-9),and apoptosis-related proteins(B-cell lymphoma-related X protein[Bax]and cleaved caspase 1)were increased to different extents after common carotid artery clipping.Among them,the expression of VCAM-1 and eNOS peaked 3 d after clipping,and the expression of endothelin-1 showed an upward trend from 1 d to 7 d after clipping.Conclusion A blunt common carotid artery trauma mouse model created by common carotid artery clipping has been successfully established.Common carotid artery clipping can induce vascular tissue injury,inflammation and cellular apoptosis,and it may be an important factor of traumatic vascular stenosis and aneurysm.
作者
李翯
刘沛
刘鹏
阳源
花伟龙
张永鑫
张磊
李子付
杨鹏飞
刘建民
LI He;LIU Pei;LIU Peng;YANG Yuan;HUA Wei-long;ZHANG Yong-xin;ZHANG Lei;LI Zi-fu;YANG Peng-fei;LIU Jian-min(Stroke Center,Changhai Hospital,Naval Medical University(Second Military Medical University),Shanghai 200433,China;The First Student Team,College of Basic Medical Sciences,Naval Medical University(Second Military Medical University),Shanghai 200433,China;The Second Student Team,College of Basic Medical Sciences,Naval Medical University(Second Military Medical University),Shanghai 200433,China)
出处
《第二军医大学学报》
CAS
CSCD
北大核心
2021年第2期127-132,共6页
Academic Journal of Second Military Medical University
基金
上海市卫生系统优秀人才培养计划(2017YQ034).
关键词
钝性创伤
颈动脉钳夹伤
内皮损伤
炎症
blunt trauma
carotid artery clipping trauma
endothelial injury
inflammatory