保护薇乔缝线携载反义寡聚核苷酸的实验研究

Study on delivering antisense oligonucleotide by Coated Vicryl

目的：探讨手术缝线携载反义基因片断的可能性及可行性，为术中血管吻合口局部转染反义基因提供新的途径。方法：保护薇乔缝线反复浸于反义核酸溶液中７２ ｈ，溴化乙锭（ＥＢ）标记，紫外灯下观察其吸附反义基因的情况。将吸附反义基因的缝线置于生理盐水中分别于不同时间点测其释放的Ｄ值，绘出体外控释曲线。同时取释放液进行电泳分析。以在荧光标记的核酸溶液中浸过的缝线直接行兔颈总动脉吻合，２４ ｈ 取材制片荧光显微镜下观察。结果：吸附核酸的保护薇乔缝线经ＥＢ标记后在紫外灯下呈烧红的铁丝样，释放曲线提示２ ｈ 释放明显增加，３６ｈ 释放达高峰。电泳则可见随时间延长而逐渐变亮的电泳带。荧光显微镜下见吻合口附近血管壁全层散在荧光分布。结论：保护薇乔缝线可较大量携载反义基因片段，可于术中血管吻合时直接将反义基因转移至吻合口并缓慢释放。

Objective:To testify the possibility and feasibility of delivering antisense oligonucleotide by thread (Coated Vicryl),to provide a new approach to antisense gene local delivery to vascular anastomosis.Methods:The Coated Vicryl was soaked in antisense oligonucleotide solution repeatedly for 72 h then marked with EB solution and observed under ultraviolet lamp.The thread carrying antisense gene emerged in normal saline and the value of optical density ( D ) at different time point,then the controlled release curve was made.Electrophoresis was also made with the released solution.The common carotid artery of rabbit was cut and anastomosed with thread soaked in fluorescein labeled antisense oligonucleotide solution,the specimen was harvested 24 hours postoperation.Results:The Coated Vicryl carrying antisense oligonucleotide marked by EB was like a reddish iron wire,the releasing curve showed the releasing increased significantly at 2.0 hour,and the releasing peak was 36 hour postoperation. Electrophoresis indicated an increasing light band.Fluorescence microscope saw the scattered fluorescent staining in the panmura of artery near the anastomosis.Conclusion:The Coated Vicryl can largly carry antisense oligonucleotide and directly deliver it to anastomosis by slow releasing.