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原位获取内皮细胞构建兔颈静脉去内皮细胞及血栓形成模型
In situ harvest of endothelial cells to develop a rabbit model of endothelial denudation and thrombosis of jugular vein

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DOI:
作者:
孙彦隽
SUN Yan-jun
作者单位:
上海交通大学医学院附属上海儿童医学中心
Shanghai Children’s Medical Center, School of Medicine, Shanghai Jiaotong University
关键词:
兔；颈静脉；内皮剥脱；原位获取；血栓形成
rabbit; jugular vein; endothelial denudation; in situ harvest; thrombosis
摘要:
摘要： 目的 通过使用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及血栓形成模型。 方法 健康新西兰实验用大白兔12只，雌雄不据，随机分为2组，体质量2.2～2.6（2.42±0.14）kg。分离兔右侧颈静脉，保留对侧，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。术后，抗凝组兔给与低分子肝素钠，对照组不给抗凝血药物，连续观察7天后，处死动物，检查右颈静脉的通畅情况。 结果 证实原位消化所分离的颈静脉细胞为内皮细胞。抗凝组1只兔术后第3天因出血死亡，尸检其右侧颈静脉通畅。其余5只存活，处死后探察右侧颈静脉，显示2只兔颈静脉通畅，1只兔颈静脉内少量血栓形成，管腔轻度狭窄，1只兔颈静脉内中等程度血栓形成，管腔中度狭窄，1只兔颈静脉完全堵塞，明显血栓形成。对照组1只兔术后第2天出现上腔静脉梗阻症状，术后第4天死亡，尸检发现右侧颈静脉和上腔静脉内血栓形成，血管腔完全堵塞。其余5只实验兔均存活，处死探察发现右侧颈静脉内均血栓形成，管腔完全堵塞。 结论 原位消化法构建兔颈静脉去内皮及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。
Abstract Objective: Harvest endothelial cells from jugular vein of rabbit in situ to develop a model of endothelial denudation and thrombosis of jugular vein. Methods: Twelve New Zealand white rabbits, bodyweight ranged from 2.2～2.6（2.42±0.14）kg, randomized into 2 groups. Right jugular vein of rabbit was dissected and kept in situ, the contralateral 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 harvest, Low molecular weight heparin sodium was only administered for rabbit of one group, no anticoagulant was administered for the other group as control. All rabbits were killed 7 days after endothelial cells harvest and patency of right jugular veins was checked. Results: Those cells harvested in situ from rabbit jugular vein were confirmed as endothelial cells. One rabbit of anti-coagulation group died of bleeding 3 days after harvest, its right jugular vein was discovered patent by autopsy. The rest 5 survived and killed 7 days after harvest, there were 2 rabbit with patent right jugular vein, no thrombosis, 1 with mild stenosis and thrombosis, 1 with moderate stenosis and thrombosis 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. it was discovered that right jugular veins were obstructed and with thrombosis among all the other 5 rabbits survived, and killed 7 days after harvest in situ. Conclusion: The method of in situ harvest endothelial cells from the jugular vein of rabbit to develop model of endothelial denudation and thrombosis of jugular vein was viable and could be use as an experimental subject for anti-coagulation, antithrombotic and thrombolytic therapy.

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