中国神经再生研究(英文版)

中国神经再生研究(英文版) ›› 2021, Vol. 16 ›› Issue (9): 1882-1889.doi: 10.4103/1673-5374.306090

• 原著:周围神经损伤修复保护与再生 • 上一篇    下一篇

质粒pBud-coVEGF165-coFGF2局部注入大鼠正常坐骨神经可诱导神经再生

  

  • 出版日期:2021-09-15 发布日期:2021-02-05
  • 基金资助:

    这项研究得到了喀山联邦大学俄罗斯政府的竞争性增长计划的支持； 俄罗斯联邦教育和科学部的国家任务，以及俄罗斯联邦总统的支持，以支持俄罗斯主要科学学校的国家支持；这项工作还得到了喀山联邦大学俄罗斯政府竞争性发展计划的支持；俄罗斯基础研究基金会支持

Angiogenesis and nerve regeneration induced by local administration of plasmid pBud-coVEGF165-coFGF2 into the intact rat sciatic nerve

Ruslan Masgutov1, 2, *, Alina Zeinalova1, Alexey Bogov2, Galina Masgutova1, Ilnur Salafutdinov1, Ekaterina Garanina1, Valeriia Syromiatnikova1, Kamilla Idrisova1, Adelya Mullakhmetova1, Dina Andreeva1, Liliya Mukhametova1, Adilet Kadyrov3, Igor Pankov3, Albert Rizvanov1#br#   

  1. 1OpenLab “Gene and Cell Technologies”, Institute of Fundamental Medicine and Biology, Kazan Federal University, Kazan, Russia; 2Republican Clinical Hospital, Kazan, Russia; 3Department of Traumatology and Orthopedics, Kazan State Medical Academy, Kazan, Russia
  • Online:2021-09-15 Published:2021-02-05
  • Contact: Ruslan Masgutov, MD, PhD, masgut@gmail.com.
  • Supported by:
    his study was supported by the Russian Government Program of Competitive Growth of Kazan Federal University; state assignment 20.5175.2017/6.7 of the Ministry of Education and Science of Russian Federation and Grant of the President of the Russian Federation for state support of the leading scientific schools of the Russian Federation. The work was also supported by the Russian Government Program of Competitive Growth of the Kazan Federal University (to AR); the Russian Foundation for Basic Research grant 18-54-45023 Ind_a (to IS and GM).

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摘要：

血管内皮生长因子（VEGF）和成纤维细胞生长因子2（FGF2）作为神经营养因子，可促进多种神经组织的再生。实验观察了大鼠正常坐骨神经神经外膜注入质粒构建体рBud-coVEGF165-coFGF2（30 ng/µL）对神经再生和血管生成的影响。(1)坐骨神经的组织学检查和血清的多重免疫测定结果显示，质粒рBud-coVEGF165-coFGF2生物安全性较好。质粒pBud-coVEGF165-coFGF2局部给药可使外周血中促炎性细胞因子肿瘤坏死因子α和白细胞介素12水平明显降低，而使以下细胞因子和趋化因子的水平升高：调节活化正常T细胞表达与分泌的趋化因子（RANTES）、表皮生长因子、白细胞介素2和单核细胞趋化蛋白1。外周血中的这些变化表明质粒具有系统作用，并可能调节免疫应答。(2) RT-PCR揭示了coFGF2，coVEGF165，ratFGF2和ratVEGFA转录物从坐骨神经的远端直接转移到近端。(3)免疫组织化学染色显示，至注射后14天，坐骨神经中VEGFA表达仍存在。(4)这些结果表明，以局部施用质粒构建体рBud-coVEGF165-coFGF2可促坐骨神经再生和血管生成。

https://orcid.org/0000-0003-2245-2182 (Ruslan Masgutov)

Abstract: Vascular endothelial growth factor (VEGF) and fibroblast growth factor 2 (FGF2) are well-known growth factors involved in the regeneration of various tissues and organs, including peripheral nerve system. In the present study, we elucidated the local and systemic effects of plasmid construct рBud-coVEGF165-coFGF2 injected into the epineurium of intact rat sciatic nerve. Results of histological examination of sciatic nerve and multiplex immunoassays of serum showed the absence of immunogenicity and biosafety of plasmid рBud-coVEGF165-coFGF2. Moreover, local administration of plasmid DNA construct resulted in significantly decreased levels of pro-inflammatory cytokines in the peripheral blood, including tumor necrosis factor α (TNFα) and interleukin-12, and significantly increased levels of cytokines and chemokines including Regulated upon Activation, Normal T Cell Expressed and Presumably Secrete (RANTES), epidermal growth factor, interleukin-2, and monocyte chemoattractant protein 1. These changes in the peripheral blood on day 7 after injection of plasmid construct рBud-coVEGF165-coFGF2 show that the plasmid construct has systemic effects and may modulate immune response. At the same time, reverse transcription-polymerase chain reaction revealed transient expression of coFGF2, coVEGF165, ratFGF2 and ratVEGFA with direct transport of transcripts from distal part to proximal part of the sciatic nerve.  Immunohistochemical staining revealed prolonged presence of VEGFA in sciatic nerve till 14 days post-injection. These findings suggest that local administration of plasmid construct рBud-coVEGF165-coFGF2 at a concentration of 30 ng/µL results in the formation of pro-angiogenic stimuli and, and the plasmid construct, used as a drug for gene therapy, might potentially facilitate regeneration of the sciatic nerve. The study was approved by the Animal Ethics Committee of Kazan Federal University, procedures were approved by the Local Ethics Committee (approval No. 5) on May 27, 2014.

Key words: chemokines, cytokines, fibroblast growth factor, gene therapy, growth factors, peripheral nerve system, sciatic nerve regeneration, vascular endothelial growth factor