上海交通大学学报（医学版）

上海交通大学学报（医学版） ›› 2019, Vol. 39 ›› Issue (7): 730-.doi: 10.3969/j.issn.1674-8115.2019.07.007

• 论著·基础研究 • 上一篇    下一篇

Pluronic F-127-胰蛋白酶混合凝胶提高静脉桥血管腺相关病毒感染效率的研究

李送 1，钟昌明 1，王小文 1, 2，张诚 1, 3，冯波 2，谢晓 4，黄春 1，向小勇 1   

  1. 1. 重庆医科大学附属第一医院心胸外科，重庆 400016；2. 香港中文大学生物医学研究院再生医学重点实验室，香港 999077；3.伦敦大学玛丽皇后学院威廉哈维研究所临床药理学中心，伦敦 EC1M 6BQ；4. 上海交通大学医学院附属新华医院心胸外科，上海 200092
  • 出版日期:2019-07-28 发布日期:2019-08-26
  • 通讯作者: 王小文，电子信箱：xiaowenwang@yeah.net。向小勇，电子信箱：cqmusurgery@163.com。为共同通信作者。
  • 作者简介:李送（1984—），男，硕士生；电子信箱： lijaden@sina.com。
  • 基金资助:
    国家自然科学基金（81700320）；重庆市科学技术委员会基础科研项目（CSTC 2015JCYA10099）

Improved efficiency of adeno-associated virus mediated gene transfer to vein graftsof pluronic F-127-trypsin gel

LI Song1, ZHONG Chang-ming1, WANG Xiao-wen1, 2, ZHANG Cheng1, 3, FENG Bo2, XIE Xiao4, HUANG Chun1, XIANG Xiao-yong1   

  1. 1. Department of Cardiothoracic Surgery, The First Affiliated Hospital of Chongqing Medical University, Chongqing 400016, China; 2. Key Laboratory for Regenerative Medicine, School of Biomedical Sciences, The Chinese University of Hong Kong, Hong Kong 999077, China; 3. Centre for Clinical Pharmacology, William Harvey Research Institute, Queen Mary University of London, London EC1M 6BQ, United Kingdom; 4. Department of Cardiovascular Surgery, Xinhua Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai 200092, China
  • Online:2019-07-28 Published:2019-08-26
  • Supported by:
    National Natural Science Foundation of China, 81700320; Chongqing Science and Technology Commission Basic Research Funding, CSTC 2015JCYA10099

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摘要： 目的 ·探讨应用 pluronic F-127-胰蛋白酶混合凝胶是否可以有效提高静脉桥血管中腺相关病毒（ adeno-associated virus，AAV）的感染效率。方法 ·构建大鼠静脉桥血管再狭窄模型。用编码增强型绿色荧光蛋白（ enhanced green fluorescent protein，Egfp）报告基因的重组 AAV（AAV1、AAV6、AAV8、AAV9血清型）感染静脉桥血管，并使用 pluronic F-127-胰蛋白酶混合凝胶增加病毒接触时间和改善血管壁通透性。分别在术后 21 d采集标本，应用冰冻切片、免疫组织化学染色和定量反转录 PCR检测静脉桥血管中 EGFP的表达以明确感染效率。通过测量组织拉伸强度来评估组织的结构完整性。结果 · Pluronic F-127-胰蛋白酶混合凝胶能够显著提高静脉桥血管中 AAV的感染效率，但并不影响静脉桥血管的组织抗拉性，且 AAV6血清型相比 AAV1、AAV8、AAV9血清型感染静脉桥血管的效率更高。结论 · Pluronic F-127-胰蛋白酶混合凝胶涂染是一种提高静脉桥血管 AAV感染效率安全有效的方法，可用于静脉桥血管再狭窄的防治研究。

关键词: 基因治疗, 腺相关病毒, pluronic F-127, 静脉桥血管, 再狭窄

Abstract:

Objective · To investigate whether pluronic F-127-trypsin gel enhances the efficiency of adeno-associated virus (AAV) mediated gene transfer to vein grafts. Methods · Rat model of venous bridge vascular restenosis was established. The vein grafts were infected with recombinant AAV encoding an enhanced green fluorescent protein (Egfp) reporter gene, and pluronic F-127-trypsin gel was used to increase viral contact time and viral penetration. The of EGFP in vein grafts was determinedfrozen section, immunohistochemistry staining and quantitative reverse transcriptase-mediated PCR 21 days after surgery. Structural integrity of the tissue was evaluatedmeasurement of tissue tensile strength. Results · Pluronic F-127-trypsin gel can significantly improve the efficiency of AAV infection in vein grafts, in which AAV serotype 6 was more efficient than AAV 1, 8, and 9 serotypes in infecting vein grafts, and tissue tensile strength was not affected. Conclusion · Pluronic F-127 trypsin gel may be a safe and effective method to improve the efficiency of AAV mediated gene transfer to vein grafts. It can be used for the prevention and treatment of venous bridge vascular restenosis.

Key words: gene therapy, adeno-associated virus, pluronic F-127, vein grafts, restenosis

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