原位获取内皮细胞构建兔颈静脉去内皮细胞和血栓形成模型  

In situ harvest of endothelial cells for developing animal model of endothelial denudation and thrombosis of jugular vein in rabbits

作　　者：孙彦隽[1] 朱丽敏[1] 徐卓明[1] 刘锦纷[1] 

机构地区：[1]上海交通大学医学院附属上海儿童医学中心心胸外科,上海200127

出　　处：《上海医学》2013年第12期1024-1027,共4页Shanghai Medical Journal

摘　　要：目的采用Ⅰ型胶原酶原位灌注消化法原位获取兔颈静脉的内皮细胞,构建兔颈静脉的内皮剥脱和血栓形成模型。方法新西兰大白兔12只,雌雄不拘,随机分入抗凝组和对照组。①分离兔右侧颈静脉,保留对侧颈静脉,采用改进的Ⅰ型胶原酶原位灌注消化法分离兔颈静脉内皮细胞,体外培养,并进行血管内皮细胞鉴定。②抗凝组兔腹部皮下注射低分子肝素钠,对照组不给予抗凝血药物,7d后处死动物,检查右颈静脉的通畅情况。结果抗凝组中,1只兔在术后第3天出现口腔和鼻腔出血,粪便带血,手术切口出血而死亡,其右侧颈静脉通畅,颈部切口内有明显血肿。存活5只:颈静脉通畅2只;颈静脉内少量血栓形成1只,管腔轻度狭窄,管腔残余内径为对侧颈静脉的80%;颈静脉内中等程度血栓形成1只,管腔中度狭窄,管腔残余内径为对侧颈静脉的50%;兔颈静脉内血栓形成、管腔完全堵塞1只。对照组中,术后第2天开始出现球结膜水肿1只,肢体和躯干运动减少,术后第4天死亡,其右侧颈静脉内血栓形成,管腔完全堵塞,血栓向下延伸到上腔静脉,造成上腔静脉管腔堵塞;右侧颈静脉内血栓形成、管腔完全堵塞5只,均存活。结论Ⅰ型胶原酶原位灌注消化法构建兔颈静脉去内皮和血栓形成模型具有一定的可行性,可为研究颈静脉血管去内皮后如何预防血栓形成的药物学方法和血栓形成以后的治疗手段提供实验对象。Objective To develop an animal model of endothelial denudation and thrombosis of jugular vein by harvesting endothelial cells from jugular vein of rabbit in situ. Methods Twelve New Zealand white rabbits were randomized into two groups. Right jugular vein was dissected and kept in situ and the left jugular vein was kept untouched and intact. Endothelial cells were detached by in situ cannulation and perfusion with type I collagenase. Harvested endothelial cells were cultured in vitro and identified by inverted microscope, transmission electron microscope and immunohistochemistry. After endothelial cells were harvested, low molecular weight heparin sodium was subcutaneously injected in the abdomen of rabbits in anticoagulation group, while no anticoagulant was administered in control group. All rabbits were killed 7 days later and right jugular veins were checked. Results One rabbit of anticoagulation group died of bleeding 3 days after harvest and its right jugular vein was discovered patent by autopsy. In the rest 5 survivals, there were 2 rabbits with patent right jugular vein and without thrombosis, 1 with mild stenosis and thrombosis （internal diameter of vein was 80% of the left one）, 1 with moderate stenosis and thrombosis （internal diameter of vein was 50% of the left one） and 1 with complete obstruction and thrombosis. One rabbit of control group died 4 days after harvest in situ of superior vena cava syndrome and thrombosis in right jugular vein and superior vena cava, confirmed by autopsy. Complete obstruction and thrombosis were found in the right jugular veins of the other 5 survival rabbits. Conclusion /n situ harvest endothelial cells from the jugular vein of rabbit is available for developing animal model of endothelial denudationand thrombosis of jugular vein, which can be used as an experimental subject for anticoagulation, antithromboticand thrombolytic therapy.

关 键 词：兔 颈静脉 内皮剥脱 原位获取 血栓形成 

分 类 号：R-332[医药卫生]