神经药理学报

神经药理学报 ›› 2021, Vol. 11 ›› Issue (2): 21-25.DOI: 10.3969/j.issn.2095-1396.2021.02.005

• 实验方法学 • 上一篇    下一篇

大鼠脊髓损伤模型的制备与评估

钱麒炳，王金瑛，赵晨宇，等   

  1. 河北北方学院基础医学院，张家口，075000，中国

  • 出版日期:2021-04-26 发布日期:2021-04-26
  • 通讯作者: 宋小青，女，副教授；研究方向：细胞生物学；E-mail：songxiaoqing0417@163.com
  • 作者简介:钱麒炳，男，临床医学专业；E-mail：yxyqqb@163.com
  • 基金资助:

    张家口市科技局应用基础研究专项（No.1911020D），河北省大学生创新创业项目（No.S202110092005），河北北方学院一般项目（No.YB2018034），河北北方学院省级重点形态学实验室支撑项目（No.XTZX201901）

Establishment and Evaluation of the Spinal Cord Injury Model in Rats

QIAN Qi-bing，WANG Jin-ying，ZHAO Chen-yu，et al   

  1. Basic Medical College of Hebei North University，Zhangjiakou，075000，China

  • Online:2021-04-26 Published:2021-04-26
  • Contact: 宋小青，女，副教授；研究方向：细胞生物学；E-mail：songxiaoqing0417@163.com
  • About author:钱麒炳，男，临床医学专业；E-mail：yxyqqb@163.com
  • Supported by:

    张家口市科技局应用基础研究专项（No.1911020D），河北省大学生创新创业项目（No.S202110092005），河北北方学院一般项目（No.YB2018034），河北北方学院省级重点形态学实验室支撑项目（No.XTZX201901）

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摘要：

目的：制备大鼠急性脊髓损伤（spinal cord injury，SCI）模型并对模型进行评估，为探究SCI 继发性反应的发生机制及SCI 早期干预治疗奠定基础。方法：将10 周龄的雄性SD 大鼠随机分为SCI 组和假手术（Sham）组（n=12），SCI 组行第9~11 胸椎（T9-11）椎板切除术，于T10 处用标定力为50 g 的动脉瘤夹完全钳夹脊髓并维持40 s，以制备急性SCI 大鼠模型。Sham 组大鼠仅行T9-11 椎板切除手术，而不损伤脊髓组织。SCI 后，根据BBB评分标准对实验动物后肢运动功能进行评价，采用HE 染色和TUNEL 染色检测实验动物脊髓组织的病理学变化和细胞凋亡，并采用 Western blot 对细胞凋亡密切相关蛋白：B淋巴细胞瘤-2（ B-cell lymphoma-2，Bcl-2）、Bcl-2相关X蛋白（Bcl-2 associated X protein，Bax）和cleaved caspase3（c.caspase3）的相对含量进行检测。结果：

SCI 组大鼠术后运动功能BBB 评分显著低于Sham 组；HE 染色显示SCI 组动物脊髓结构破坏严重，细胞胞体收缩，细胞间隙增大，灰质部位空洞明显；而Sham 组未见明显异常。TUNEL 染色显示SCI 组脊髓组织的细胞凋亡率显著高于Sham 组；Western blot 结果显示，SCI 可致脊髓组织抗凋亡蛋白Bcl-2 的相对量降低，却使促细胞凋亡蛋白Bax 和细胞凋亡效应酶c.caspase3 的相对含量升高。结论：利用动脉瘤夹压迫法制备大鼠急性SCI 模型，实验操作简单易行、稳定性良好，在SCI 修复研究中值得推广应用。

关键词:

font-family:Helvetica, "> 脊髓损伤, 模型制备, font-family:Times, ">BBB 评分, 细胞凋亡

Abstract:

Objective：To prepare and evaluate the model of acute spinal cord injury（SCI）in rats，so as to lay a foundation for exploring the mechanism of SCI secondary reaction and the early intervention of SCI repair. Methods：The male SD rats aged 10 weeks were randomly divided as SCI group and Sham group（n=12）. In SCI group，the thoracic vertebrae laminectomy was performed at the 9-11th thoracic vertebrae（T9-11）. At T10，the spinal cord was completely clamped with an aneurysm clamp（50 g for 40 s），to prepare the acute SCI model in rats. The rats in Sham group only underwent laminectomy at T9-11. After SCI，the hindlimb motor function of rats was evaluated according to BBB scores. HE staining and TUNEL staining were respectively conducted to detect the pathological changes and cell apoptosis of spinal cord tissue. The relative content of proteins closely related to cell apoptosis was detected by Western blot. Results：The BBB scores of motor function in SCI group were significantly lower than those in Sham group；HE staining showed that the spinal cord structure of SCI group was severely damaged，cell body contracted，intercellular space became increased，and gray matter cavities were obviously seen；There was no abnormal change in Sham group. TUNEL staining demonstrated that the rate of cell apoptosis in SCI group was significantly higher than that of Sham group；Western blot results showed that SCI significantly decreased the relative content of Bcl-2，but increased the relative content of Bax and caspase-3. Conclusion：The SCI model prepared with an aneurysm clip compression could reflect the pathophysiology of the injured spinal cords. In view of the stability and simplicity of the model making，the clip compression should be widely recommended in the research of SCI repair.

Key words:

font-family:Times, "> spinal cord injuryfont-family:Helvetica, ">, model makingfont-family:Helvetica, ">, BBB scorefont-family:Helvetica, ">, cell apoptosis

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