应用生物素葡聚糖胺标记脊髓损伤后神经再生动物模型的建立

Establishment of nerve regneration animal model following spinal cord injury labed with biotinylated dextran amine

目的应用立体定向技术注射生物素葡聚糖胺(BDA)标记脊髓损伤后微管亲合力调节激酶2(MARK2)在神经再生中作用的动物模型。方法SD大鼠16只,随机分为正常对照组(4只)和脊髓损伤组(12只),脊髓损伤组采用剪刀离断脊髓,12只大鼠建立SCI模型后,应用立体定向技术,分别注射生理盐水(4只),增强型绿色荧光蛋白(EGFP)-腺病毒(4只),MARK2-腺病毒(4只),1周后16只大鼠均在皮层运动区注射BDA示踪剂,2周后处死大鼠,取出全脑和脊髓。结果脊髓损伤组大鼠双下肢均截瘫,肌力0级(成功率为100%),在皮层运动区立体定向注射BDA后,行激光共聚焦检查BDA所标记的神经传导束,检出率为94%。结论此种方法建立的动物模型安全可靠,重复性良好。

Objective To study the role of microtubule affinity- regulating kinase 2 (MARK2) in nerve regeneration in an animal model after spinal cord injury (SCI) labeled with biotinylated dextran amine (BDA) using stereotactic injection of. Methods Sixteen SD rats were randomly divided into control group (4 rats) and SCI groups (12 rats). Spinal cord was cut off by scissors in spinal cord injury group. After SCI model was established in 12 rats, normal saline subgroup (4 rats), EGFP-adenovirus subgroup (4 rats), and MARK2-adenovirus subgroup (4 rats) were given the injection of normal saline, EGFP-adenovirus and MARK2-adenovirus into cortical motor area respectively by stereotactic injection technique. One week later, 16 rats were injected with BDA in cortical motor area. After 2 weeks, the animals were executed and the whole brain and spinal cord were removed. Results In SCI groups, rats had lower limb paraplegia (100% success rate). After stereotactic injection of BDA into the motor cortex, the nerve conduction tracts labeled by BDA were confirmed by confocal laser scanning (94% success rate). Conclusion The animal model established by this method is safe, reliable and reproducible.