摘要
目的探讨大鼠脊髓损伤(spinal cord impairment,SCI)后Nogo受体拮抗剂NEP1-40对降钙素基因相关肽(calcitoningene related peptide,CGRP)表达的影响。方法取雌性SD大鼠,随机分成对照组、生理盐水组和NEP1-40治疗组,每组60只。生理盐水组和NEP1-40治疗组的大鼠建立脊髓半切损伤模型,并在蛛网膜下腔留管,再分别注射生理盐水和NEP1-40;对照组的大鼠不损伤脊髓。SCI后1、3、7、14、21 d收集损伤部位的脊髓组织,再用免疫荧光、免疫印迹检测每组CGRP蛋白含量,并在术后7 d采用免疫荧光双标技术检测损伤部位的脊髓内生长相关蛋白-43(growth associated protein 43,GAP-43)、CGRP的表达量。结果生理盐水组CGRP阳性神经细胞减少,而NEP1-40组CGRP阳性神经细胞24 h内减少,然后开始增多,到术后7 d达到高峰,之后一直维持在高水平表达,而对照组发现CGRP阳性神经细胞在低水平表达。免疫印迹和免疫荧光的结果一致。术后7 d,NEP1-40组GAP-43阳性神经细胞数最多,生理盐水组次之,对照组未发现,而且发现GAP-43和CGRP有共表达。结论脊髓损伤后,NEP1-40可以降低神经细胞Nogo-A的表达而大量增加CGRP、GAP-43的表达,从而改善脊髓的内环境,促进神经突起的修复。
The study designed to investigate the effect of NEP1-40 on calcitonin gene related peptide （CGRP） expression after spinal cord impairment （SCI） in rats. Adult female SD rats were randomly divided into three groups： NEP1-40 group, normal saline （NS） group and sham group. In the NEP1-40 and NS group, the SCI model was produced by the spinal cord hemiseetion, while the rats in sham group received sham operation without the spinal hemisection. NEP1-40 and NS groups were intrathecally injected with NEP1-40 and normal saline, respectively. At different time points after SCI, injured region of spinal cord was taken out as sample. The levels of CGRP expression were measured by immunofluorescence technique and Western blot. And the expressions of growth associated protein 43 （GAP-43） and CGRP were detected using double immunofluorescent staining. Compared with sham group, the CGRP expression in NS group was significantly down-regulated during the whole time after SCI. In NEP1-40 group, the level of CGRP expression was down-regulated on 1 d after SCI, then up-regulated on 3 to 21 d after SCI. Western blot showed an identical result to that of immunofluorescent staining. The double labeling results showed that on 7 d after SCI, the number of GAP-43 positive cells in NS group was lower than that in NEP1-40 group, but higher than that in sham groups. Moreover, GAP-43 and CGRP were detected in NS groups and NEPl- 40 group. These results suggest that NEP1-40 can decrease the expression level of Nogo-A, but increase the expression level of CGRP or GAP-43 after SCI, thus improving microenvironment of the injured region and promoting axonal growth and extension.
出处
《免疫学杂志》
CAS
CSCD
北大核心
2013年第8期654-658,共5页
Immunological Journal
基金
河南省教育厅科学技术研究重点项目(13B310163)
