联合应用免疫抑制剂治疗大鼠急性脊髓损伤效果观察

Experimental study of combined application of immunosuppressant agents on acute spinal cord injury in rats

目的探讨联合应用免疫抑制剂他克莫司(FK506)和FTY720对大鼠急性脊髓损伤的神经保护作用。方法采用Allen′s打击法制备大鼠脊髓损伤模型,84只大鼠分成4组:FK506+FTY720联合治疗组(A组)、FK506治疗组(B组),脊髓损伤组(C组)和正常对照组(D组)。A组通过尾静脉注射0.3 mg/kg FK506,并予鼻饲0.3 mg/kg FTY720,B组通过尾静脉注射0.5 mg/kg FK506,C组注射生理盐水。伤后6 h、24 h和48 h取伤段脊髓行HE染色分析和Tunel凋亡分析;伤后第1、3和7天行体感诱发电位(SEP)检测;伤后第1、3、7和14天行BBB评分和斜板试验评分。结果联合治疗组和FK506治疗组伤后各个时间取材点的脊髓损伤出血坏死区均轻于损伤组,在第6、24和48小时时间点脊髓Tunel神经细胞凋亡检测明显轻于损伤组,有显著差异性(P<0.05),但2组之间比较无显著差异性(P>0.05)。联合治疗组和FK506治疗组伤后各时间点SEP、BBB评分和斜板试验评分均优于损伤组,有显著性差异(P<0.05),但是2组之间差异无统计学意义(P>0.05)。结论大鼠急性脊髓损伤早期联合应用FK506和FTY720具有良好的保护神经功能和促进神经功能恢复的作用。

Objective To explore the effect of combined application of immunosuppressant agents tocrolimus and FTY720 on acute spinal cord injury （SCI） in rats. Methods Eight-four male rats were randomly divided into four groups： the combination therapy group （group A）, the FK506 treatment group （group B）, the injury group （group C） and the control group （group D）. The rats of groups A, B and C were subjected SCI by Allen＇s model, and group D just did laminectomy. The rats of group A were injected with FK506 0.3 mg/kg through caudal vein and nasal feeding with FTY720 0.3 mg/kg, while group B was injected with FK506 0.5 mg/kg and C was injected with saline. H＆E analysis and apoptosis of neural cells analysis of the injured spinal cord were conducted at 6 h, 24 h and 48 h. SEP analysis was on days I, 3 and 7. Neurological function was used on day 1, 3, 7 and 14. Results H＆E stain showed that hemorrhage and necrosis in injured spinal cord of the combination therapy group and FK506-treatment group were less serious than those in the injury group in all time after SCI. Tunel analysis demonstrated the mount of positive cells in the combina- tion therapy group and FKS06-treatment group were less than the injury group in all time after SCI （P 〈 0.05）, but there was no significant difference between the two groups （P 〉 0.05）. The re- sults of SEP, BBB score and inclined plate test score at each time in the combination therapy group and FK506. Conclusion The combined application of FK506 and FTY72 on acute spinal cord injury in rats can well protect the neurological function and promote the neurological recovery.