解剖学报

解剖学报 ›› 2021, Vol. 52 ›› Issue (4): 657-661.doi: 10.16098/j.issn.0529-1356.2021.04.024

• 技术方法 • 上一篇    下一篇

缝线法构建兔角膜新生血管模型

杜立群 孙嘉璋 米丰花 吴欣怡*   

  1. 山东大学齐鲁医院眼科，济南 250012
  • 收稿日期:2020-03-17 修回日期:2020-07-02 出版日期:2021-08-06 发布日期:2021-08-06
  • 通讯作者: 吴欣怡 E-mail:xywu8868@126.com
  • 基金资助:
    北京白求恩公益基金会

Construction of corneal neovascularization rabbit model by suture

DU Li-qun  SUN Jia-zhang  MI Feng-hua  WU Xin-yi*   

  1. Department of Ophthalmology, Qilu Hospital, Shandong University, Shandong Jinan 250012, China
  • Received:2020-03-17 Revised:2020-07-02 Online:2021-08-06 Published:2021-08-06
  • Contact: WU Xin-yi E-mail:xywu8868@126.com

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摘要：

目的  比较不同缝线及缝合方式对兔角膜新生血管（CNV）模型的诱导效果。   方法  健康新西兰大白兔 45只，应用角膜缝线法制作CNV模型，根据缝线及缝合方式的不同随机平均分成5组，裂隙灯观察CNV生长趋势并摄片记录。筛选新生血管诱导稳定有效者，于角膜缝线前、缝合后7 d和14 d分别抽取动物房水，ELISA法检测血管内皮生长因子（VEGF）蛋白含量，并于相同时间点随机处死实验兔各3只，取角膜行HE染色观察组织学变化，免疫荧光染色观察VEGF表达。   结果  8-0缝线单套环平行（A1）和垂直缝合组（A2）角膜缝线分别于8 d和14 d后脱落，血管萎缩，模型制作失败；10-0缝线双套环平行（B1）和垂直缝合组（B2）14 d时分别于角膜缘见稀疏和短簇状血管，CNV诱导效率不高。10-0缝线3套环放射状缝合组（B3）CNV生长旺盛，呈束状，房水VEGF表达较缝线前增强，14 d 高于7 d。光学显微镜检查显示缝线后7 d时，B3组角膜水肿，基质CNV。VEGF免疫荧光染色见少量着色；14 d时，B3组角膜基质CNV及VEGF免疫荧光染色均较7 d时增多。   结论  角膜缝线法可诱导兔角膜CNV模型；10-0线3套环缝合法（B3）诱导CNV稳定有效，为有效诱导方法。

关键词: 角膜, 新生血管, 缝线, 缝合, 酶联免疫吸附测定, HE染色, 免疫荧光, 兔

Abstract:

Objective  To compare the effect of different sutures and suture method  on corneal neovascularization(CNV) in rabbit models.  
  Methods  NV was induced by placing sutures at the corneal periphery of rabbits (n=45). To observe the NV status, 45 rabbits were randomly divided into 5 equal groups. Group A applied 8-0 absorbable suture (A1 single loop parallel suture, A2 single loop vertical suture). In group B, 10-0 nylon suture was used (B1 double loop parallel suture, B2 double loop vertical suture, B3 three loop radial suture). The development of CNV was observed with slit lamp microscope and photographed. Therefore the effective model for neovascularization induction was selected. Histological examination, immunofluorescent staining and ELISA analysis for the vascular endothelial growth factor(VEGF) were performed before suture, 7 and 14 days after suture.    Results  Sutures fell off and CNV gradually atrophied in group A1 and A2; At the 14th day after suture, Sparse or short cluster CNV grew into the corneal margin in group B1 and B2, while CNV was vigorous and grew in bundles in group B3. The expression of VEGF in aqueous humor increased in B3 group after suturing, and increased in 14 days as compared with 7 days after suture. Corneal edema, neovascularization and little immunofluorescence staining for VEGF were detected in group B3 after 7 days suture. More neovascularization and immunofluorescence staining for VEGF were detected in group B3 after 14 days suture.   Conclusion  Corneal NV can be induced successfully in rabbit model by suturing. The method  of 10-0 thread with three sets of circular seams(B3) is stable and effective.

Key words: Cornea, Neovascularization, Suture, Sew up, Enzyme-linked immunoassay, HE staining, Immunofluoresence, Rabbit

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