组织工程与重建外科杂志 ›› 2016, Vol. 12 ›› Issue (6): 349-352.doi: 10.3969/j.issn.1673-0364.2016.06.005

• 论著 • 上一篇    下一篇

改良小鼠后肢淋巴水肿模型的构建

孙一宇,崔春晓,戴婷婷,蒋朝华,曹卫刚,李圣利   

  1. 上海交通大学医学院附属第九人民医院整复外科
  • 发布日期:2020-07-23

An Improvement Model of Secondary Lymphedema in Hind Limb of Mouse

SUN Yiyu,CUI Chunxiao,DAI Tingting,JIANG Chaohua,CAO Weigang,LI Shengli   

  1. Department of Plastic and Reconstructive Surgery, Shanghai Ninth People’s Hospital, Shanghai Jiaotong University School of Medicine;
  • Published:2020-07-23
  • Contact: 国家自然科学基金(81501571)

摘要: 目的通过调整辐射剂量和手术时机,构建一种更为稳定和持久的小鼠后肢淋巴水肿模型。方法将36只8周龄雌性C57小鼠随机分为3组:术前放疗组(R+S)、术后放射组(S+R)、术前及术后放射组(r+S+r)。采用淋巴结摘除手术联合局部放射的方法进行模型构建。R+S组于术前3 d接受4.5 Gy的局部放射,S+R组于术后2周接受4.5 Gy的局部放射,r+S+r组于术前3 d和术后2周分别接受2.25 Gy的局部放射。术后采用排水法测量动物后肢体积变化。术后24周时在后肢注射FDNG以显示体内淋巴管的分布,并结合免疫组化计数微淋巴管数目来评价淋巴管新生的情况。结果在我们为期6个月的观察中,所有小鼠均成功形成淋巴水肿。尤以r+S+r组形成的淋巴水肿最为明显、稳定持久,且死亡率和并发症率较低。术后24周的淋巴管造影显示,R+S及S+R组后肢浅表淋巴管密度增加,r+S+r组少有淋巴管再生。免疫组化结果显示,r+S+r组的淋巴管数量要少于R+S及S+R组(P<0.05)。结论在术前3 d及术后2周分别接受2.25 Gy的局部放射,可以成功构建稳定持久的小鼠后肢淋巴水肿模型。

关键词: 淋巴水肿, 动物模型, 淋巴管, 放射

Abstract: Objective To establish a more stable and sustainable model of mouse lymphedema by adjusting the administrating timing and radiation doses. Methods Thirty-six female C57 mice aged 8weeks were randomly divided into 3 groups: "radiation before surgery" group (R+S, n=12), "radiation after surgery" group (S+R, n=12), and "radiation before and after surgery" group (r+S+r, n=12). The experimental groups received radiation at varied time points: S+R group, 4.5 Gy at 2 weeks after surgical treatment; R+S group, 4.5 Gy at 3 days before surgery; and r+S+r group, 2.25 Gy at both 3 days before and 2 weeks after surgery. The volume change of the hind limbs of animals was measured by drainage method. Lymphangiogenesis in vivo situation was visualized by FDNG injection through the hind limb at 24th week after operation and immunohistochemistry staining was performed to show superficial lymphatic vessels. Results In six-month observation, all mice in the experimental groups formed lymphedema successfully. The r+S+r group showed the most stable hind limb lymphedema with lower mortality and morbidity rates. At 6 months post operation,according to the lymphatic mapping, the lymphatic vessels in R+S and S+R group increased, while rare lymphatic regeneration was observed in the r+S+r group. Immunohistochemistry revealed a decreased number of lymphatic vessels in the r+S+r group, compared with other groups (P〈0.05). Conclusion A radiation dose of 2.25 Gy administered 3 days before surgery and 2 weeks after operation successfully enhanced the stability of the lymphedema model of mice hind limb.

Key words: Lymphedema, Animal model, Lymphatic vessels, Radiation

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