大鼠脊髓损伤模型的制备与评估  

Establishment and Evaluation of the Spinal Cord Injury Model in Rats

作　　者：钱麒炳 王金瑛 赵晨宇 郭任伟 宋小青[1] 魏会平[1] 朱登祥[1] QIAN Qi-bing;WANG Jin-ying;ZHAO Chen-yu;GUO Ren-wei;SONG Xiao-qing;WEI Hui-ping;ZHU Deng-xiang(Basic Medical College of Hebei North University,Zhangjiakou,075000,China)

机构地区：[1]河北北方学院基础医学院,张家口075000

出　　处：《神经药理学报》2021年第2期21-25,共5页Acta Neuropharmacologica

基　　金：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)。

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(T_(9-11))椎板切除术,于T_(10)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T_(9-11)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。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 9-11^(th) thoracic vertebrae(T_(9-11)).At T_(10),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 T_(9-11).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.

关 键 词：脊髓损伤 模型制备 BBB评分 细胞凋亡 

分 类 号：R683.2[医药卫生—骨科学]