芬苯达唑对大鼠脊髓损伤后免疫反应和运动功能恢复的影响

The influence of fenbendazole on immune response and motor function recovery of rats after spinal cord injury

目的观察芬苯达唑对脊髓损伤大鼠的CD45R阳性B细胞、IgG免疫反应以及运动功能恢复的影响。方法将75只成年雌性SD大鼠随机分为假手术组、模型对照组和芬苯达唑组,每组25例。假手术组、模型对照组术前4周给予常规啮齿动物饲料喂食(含18%蛋白质的饲料),芬苯达唑组术前4周给予加入芬苯达唑的常规啮齿动物饲料喂食。用Allen法构建脊髓损伤模型,分别于术后第1、7、14、21、28天行Basso-Beatti-Bresnahan(BBB)运动功能评估,利用免疫组化检测损伤部位脊髓组织中IgG的表达水平,利用免疫荧光检测脊髓损伤部位CD45R阳性B细胞的信号水平。结果脊髓损伤后,模型对照组和芬苯达唑组的BBB评分均显著低于假手术组(均P<0.05),随着时间的延长,两组的BBB运动评分均逐渐有所恢复,但仍低于假手术组;脊髓损伤后第1天,模型对照组与芬苯达唑组的BBB运动评分分别为(3.10±0.29)分、(3.23±0.48)分,差异尚无统计学意义;而在脊髓损伤后的第7、14、21、28天,芬苯达唑组的BBB评分均高于模型对照组,差异均有统计学意义(均P<0.05)。免疫组化检查证实损伤部位脊髓标本的IgG水平显著升高,第7天开始,芬苯达唑组各个时间点的IgG免疫反应水平均低于模型对照组,差异均有统计学意义(均P<0.05)。免疫荧光检查证实脊髓损伤后损伤部位脊髓节段CD45R阳性B细胞信号水平显著升高,第7天开始,芬苯达唑组各个时间点脊髓损伤部位的CD45R阳性B细胞的信号水平均低于模型对照组,差异均有统计学意义(均P<0.05)。结论芬苯达唑预处理可降低脊髓组织中IgG的表达水平及脊髓损伤部位的CD45R阳性B细胞的信号水平,促进脊髓损伤后的神经功能恢复。

Objective To investigate the effect of fenbendazole on CD45R (+) B cells, IgG immune response and movement function recovery of rats with spinal cord injury (SCI). Methods Seventy?five adultfemale SD rats were randomly divided into sham operation group, model control group and fenbendazole group,25 for each group. Routine rodent feed (containing 18% protein feed) was given in sham operation group andmodel control group for 4 weeks preoperatively, and fenbendazole and conventional rodents feed was given infenbendazole group for 4 weeks preoperatively. The SCI was produced using a modified Allen method. Basso?Beattie?Bresnahan (BBB) scale was used to estimate the neurological recovery on days 1, 7, 14, 21 and 28postoperation after SCI. The expression levels of IgG in the spinal cord tissue were evaluated using the immuno?histochemistry. Immunofluorescent assay was performed to identify the signal level of CD45R positive cells inthe injured spinal cord. Results BBB scores in model control group and fenbendazole group were significantlylower than in control group (P<0.05) after SCI. With the extension of time, BBB scores in model control groupand fenbendazole group were recovered gradually, but still lower than in control group. On the 1st day afterSCI, BBB movement scores in model control group and fenbendazole group were 3.10±0.29 and 3.23±0.48,respectively, and no statistically significant difference between two groups. On the day 7, 14, 21, 28 afterSCI, BBB scores in fenbendazole group were significantly higher than those in model control group (P<0.05). Immunohistochemistry revealed that IgG levels increased significantly after SCI. From day 7, the immune response of IgG levels in the fenbendazole group was weaker than that of model control group with the difference increased significantly after SCI. From day 7, CD45R positive signal levels in the fenbendazole group were low?er than in the model control group at each time point with the difference being statistically significant (P<0.05). Conclusion Fenbendazole pretreatment reduced the injury?induced CD45Rpositive B cell signal inten?sity and IgG immunoreactivity at the lesion epicenter after contusive SCI, and promoted neural functional recovery after SCI.