南方医科大学学报

南方医科大学学报 ›› 2021, Vol. 41 ›› Issue (11): 1686-1691.doi: 10.12122/j.issn.1673-4254.2021.11.13

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抑制转化生长因子β表达可以减少纤连蛋白的形成并促进小鼠脊髓损伤的修复

李夏林，潘大宇   

  1. 华中科技大学协和深圳医院脊柱外科，广东 深圳 518000；天津医科大学总医院骨科，天津 300052
  • 出版日期:2021-11-20 发布日期:2021-12-10

Inhibition of TGF-β promotes functional recovery of spinal cord injury in mice by reducing fibronectin deposition

LI Xialin1, PAN Dayu   

  1. Department of Spinal Surgery, Huazhong University of Science and Technology Union Shenzhen Hospital, Shenzhen 518000, China; Department of Orthopedics, Tianjin Medical University General Hospital, Tianjin 300052, China
  • Online:2021-11-20 Published:2021-12-10

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摘要： 目的 探讨抑制转化生长因子β（TGF-β）表达对小鼠脊髓损伤的修复作用。方法 将12只小鼠分为3组:治疗组、对照组和阴性对照组，4只/组，重复实验2次。治疗组通过对脊髓损伤半切模型小鼠进行腹腔注射TGF-β中和抗体（1D11）来抑制TGF-β的表达量；对照组小鼠注射载体抗体（13C4）作为对照，损伤后3次/周，每次25 μL/只；阴性对照组未做半切损伤，4周后心脏灌注取材，选取损伤部位上下1~2 cm脊髓进行包埋，损伤区域的免疫荧光染色通过FSP1观察成纤维细胞募集，通过Fibronectin来观察纤连蛋白沉积以及通过PGP9.5来观察神经学恢复情况。最后选择脊髓损伤钳夹模型来进一步验证，使用免疫荧光染色通过5-HT来观察轴突存活情况以及通过GFAP来观察星形胶质细胞分布情况。结果 在脊髓损伤半切模型中，与对照组相比，治疗组损伤区域中成纤维细胞募集减少（P<0.05），纤连蛋白沉积减少（P<0.05），神经学功能PGP9.5染色有所恢复（P<0.05）。在脊髓损伤钳夹模型中，1D11组与13C4组相比，5-HT阳性的轴突存活增加且下行距离延伸，GFAP阳性星形胶质细胞在损伤区域内分布数量增加（P<0.05）。结论 抑制脊髓损伤后TGF-β的表达量可以减少成纤维细胞的募集和纤连蛋白的沉淀，从而促进脊髓损伤后神经学功能恢复来修复脊髓损伤。

关键词: 脊髓损伤；转化生长因子β；成纤维细胞；纤连蛋白

Abstract: Objective To investigate the effect of transforming growth factor (TGF-β) inhibition on functional recovery of spinal cord injury in mice. Methods Twelve mice were divided into treatment group, control group and sham-operated group (n=4). The mice in the treatment group were subjected to hemisection of the spinal cord and received intraperitoneal injection of TGF-β neutralizing antibody (1D11) 3 times a week (25 μL each time), and those in control group were injected with the vehicle antibody (13C4) following spinal cord hemisection. The sham-operated mice underwent sham operation to expose the spinal cord without hemisection. Four weeks later, the heart of the mice was perfused and 1-2 cm of the spinal cord spanning the injury site was harvested. Immunofluorescence staining of FSP1, fibronectin, and PGP9.5 was performed to assess fibroblast recruitment in the injury area, fibronectin deposition, and neurological recovery. For further verification of the results, we used a mouse model of spinal cord clamp injury to observe the survival of axons and distribution of astrocytes by detecting expressions of 5-HT and GFAP with immunofluorescence assay. Results In the hemisection injury model, fibroblasts recruitment and fibronectin deposition in the injured area was significantly reduced and the neurological function was improved in 1D11 treatment group as compared with those in 13C4-treated group (P<0.05). In the spinal cord clamp injury model, treatment with 1D11, as compared with the 13C4, resulted in significantly increased number of 5-HT-positive axons with extended axonal length and obviously increased the number of GFAP-positive astrocytes in the injured area (P<0.05). Conclusion Inhibiting TGF-β after spinal cord injury can reduce the recruitment of fibroblasts and fibronectin deposition to promote recovery of neurological function and repair of the injured spinal cord in mice.

Key words: spinal cord injury; transforming growth factor-β; fibroblasts; fibronectin