中国神经再生研究(英文版)

中国神经再生研究(英文版) ›› 2019, Vol. 14 ›› Issue (12): 2199-2208.doi: 10.4103/1673-5374.262596

• 原著:周围神经损伤修复保护与再生 • 上一篇    

新鲜人羊膜包裹可有效促进损伤腓总神经的修复

  

  • 出版日期:2019-12-15 发布日期:2019-12-15
  • 基金资助:

    贵州省科技重大专项（Qin Ke He Zhong Da Zhuan Xiang Zi [2011]6002），省市科技特别合作基金项目（Sheng Shi He (2014) 59）

Fresh human amniotic membrane effectively promotes the repair of injured common peroneal nerve

Zhong-Yuan Zhang 1 , Jin Yang 1, 2 , Zhen-Hai Fan 1, 3 , Da-Li Wang 4 , Yu-Ying Wang 1, 3 , Tao Zhang 1, 3 , Li-Mei Yu 1, 3 , Chang-Yin Yu 5   

  1. 1 Key Laboratory of Cell Engineering in Guizhou Province, The Affiliated Hospital of Zunyi Medical University, Zunyi, Guizhou Province, China
    2 Department of Thyroid and Breast Surgery, Fifth People’s Hospital of Chengdu, Chengdu, Sichuan Province, China
    3 The Team of Scientific and Technological Innovation Talents on The Basic and Clinical Research of Amniotic Membrane and Bone Marrow Stem Cells in Guizhou Province, Zunyi, Guizhou Province, China
    4 Department of Burn and Plastic Surgery, The Affiliated Hospital of Zunyi Medical University, Zunyi, Guizhou Province, China
    5 Department of Neurology, The Affiliated Hospital of Zunyi Medical University, Zunyi, Guizhou Province, China
  • Online:2019-12-15 Published:2019-12-15
  • Contact: Li-Mei Yu, PhD, MD, ylm720@sina.com; Chang-Yin Yu, MD, yuchangyin6812@126.com.
  • Supported by:

    This study was supported by Guizhou Province Major Special Projects in Science and Technology of China, No. Qin Ke He Zhong Da Zhuan Xiang Zi [2011]6002; and the Special Co-operation Funds of the Science and Technology Administration in Provinces and Cities of China, No. Sheng Shi He (2014) 59 (both to LMY).

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摘要：

对于完全断裂的周围神经，缝合和自体移植是目前主要的治疗方法，但是微环境失衡和周围组织粘连都会影响神经再生和修复的质量。有研究显示人羊膜可促进多种组织的愈合。因而实验首先行5mm横断大鼠右侧腓总神经缺损建立损伤模型，以使用10/0不可吸收的手术线缝合，然后采用双层新鲜羊膜包裹缝合的受损腓总神经，并吻合型快速医用胶粘合羊膜边缘。(1)以步态分析检测大鼠后肢运动功能，通过神经电刺激计算腓总神经的传导速度，采用快蓝逆行荧光示踪观察腓总神经逆行轴浆运输情况，以苏木精-伊红染色检测腓总神经病理学变化，以实时PCR分析神经组织中轴突再生相关神经营养因子和抑制因子神经生长因子、神经生长相关蛋白43、脑衰反应调节蛋白2、脑源性神经营养因子、Lingo-1及神经胶质源神经营养因子的mRNA表达情况；(2)结果显示羊膜包裹的损伤神经功能明显得到快速改善，损伤侧后肢趾展功能提高，神经传导速度恢复，脊髓逆向轴浆运输的快蓝颗粒增加，促进胫骨前肌肉恢复，同时轴突再生相关神经营养因子神经生长因子、神经生长相关蛋白43、脑衰反应调节蛋白2和脑源性神经营养因子的mRNA表达水平恢复接近正常，而轴突再生相关抑制因子Lingo-1 mRNA表达明显下降，但神经胶质源神经营养因子mRNA表达未发生明显变化，且上述变化比单纯缝合的效果更为显著；(3)上述结果说明，羊膜包裹技术的应用可快速促进腓总神经损伤的修复，且其可能通过调节轴突再生相关神经营养因子和抑制因子表达水平而发挥作用。实验于2017年12月1日获得遵义医科大学附属医院研究伦理委员会批准（批准号112）。

orcid: 0000-0002-3377-1202 (Li-Mei Yu)

关键词: 人羊膜, 轴突, 许旺细胞, 吻合型快速医用胶, 神经缝合, 胫骨前肌肉, 神经生长因子, 腓总神经损伤, 神经再生

Abstract:

Suture and autologous nerve transplantation are the primary therapeutic measures for completely severed nerves. However, imbalances in the microenvironment and adhesion of surrounding tissues can affect the quality of nerve regeneration and repair. Previous studies have shown that human amniotic membrane can promote the healing of a variety of tissues. In this study, the right common peroneal nerve underwent a 5-mm transection in rats. Epineural nerve repair was performed using 10/0 non-absorbable surgical suture. The repair site was wrapped with a two-layer amniotic membrane with α-cyanoacrylate rapid medical adhesive after suture. Hindlimb motor function was assessed using footprint analysis. Conduction velocity of the common peroneal nerve was calculated by neural electrical stimulation. The retrograde axoplasmic transport of the common peroneal nerve was observed using fast blue BB salt retrograde fluorescent staining. Hematoxylin-eosin staining was used to detect the pathological changes of the common peroneal nerve sputum. The mRNA expression of axon regeneration-related neurotrophic factors and inhibitors was measured using real-time polymerase chain reaction. The results showed that the amniotic membrane significantly improved the function of the injured nerve; the toe spread function rapidly recovered, the nerve conduction velocity was restored, and the number of fast blue BB salt particles were increased in the spinal cord. The amniotic membrane also increased the recovery rate of the tibialis anterior muscle and improved the tissue structure of the muscle. Meanwhile, mRNA expression of nerve growth factor, growth associated protein-43, collapsin response mediator protein-2, and brain-derived neurotrophic factor recovered to near-normal levels, while Lingo-1 mRNA expression decreased significantly in spinal cord tissues. mRNA expression of glial-derived neurotrophic factor did not change significantly. Changes in mRNA levels were more significant in amniotic-membrane-wrapping-treated rats compared with model and nerve sutured rats. These results demonstrate that fresh amniotic membrane wrapping can promote the functional recovery of sutured common peroneal nerve via regulation of expression levels of neurotrophic factors and inhibitors associated with axonal regeneration. The study was approved by the Committee on Animal Research and Ethics at the Affiliate Hospital of Zunyi Medical University, China (approval No. 112) on December 1, 2017.

Key words: nerve regeneration, human amniotic membrane, axonal, Schwann cells, α-cyanoacrylate rapid medical adhesive, neural suture, tibial anterior muscle, neuronal growth factor, common peroneal nerve injury, neural regeneration