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目的:研究大鼠坐骨神经压榨模型的钙结合蛋白Calretinin(CR)在脊髓的时空变化规律,为探讨其在神经再生中的作用提供实验依据。方法:36只SD大鼠随机分为假手术对照组和坐骨神经压榨组,实验组压榨后分别存活1d到21d,免疫组化结合图像分析技术观察CR在脊髓分布和含量的变化。结果:在对照组,CR样阳性神经元主要分布于腰髓背角Ⅰ,Ⅱ层,Ⅲ~Ⅵ层只观察到一些散在分布的CR样阳性神经元,脊髓前角Ⅷ层和Ⅸ层内也可见一些多极的中间型阳性神经元。坐骨神经压榨1d后,分布于腰髓背角Ⅱ层内的CR样阳性神经元比对照组有轻微增加。3d后,CR样阳性神经元与对照组相比没有明显改变。7d后,CR样阳性神经元有轻微的减少;14d后,CR的表达显著下降;至21d,CR的表达有所恢复,但仍低于7d组。脊髓后角CR免疫阳性产物灰度值测定结果显示:术后14d后角CR表达最低,与对侧和对照组相比有统计学意义(P<0.05)。结论:坐骨神经压榨后CR表达变化呈现一定的时空模式,为进一步揭示CR在神经系统疾病中的作用提供实验依据。
OBJECTIVE: To investigate the spatial and temporal changes of Calretinin (CR), a calcium channel-binding protein in rat sciatic nerve crush model, to provide an experimental basis for exploring its role in nerve regeneration. Methods: Thirty-six Sprague-Dawley rats were randomly divided into sham operation group and sciatic nerve compression group. The rats in the experimental group survived for 1 day to 21 days after compression. The distribution and content of CR in the spinal cord were observed by immunohistochemistry and image analysis techniques. Results: In the control group, CR-positive neurons mainly distributed in the spinal cord dorsal horn Ⅰ, Ⅱ, Ⅲ ~ Ⅵ layer only observed some scattered CR-like positive neurons, spinal cord anterior chamber Ⅷ and Ⅸ layer also Some multipolar positive neurons can be seen. One day after sciatic nerve crushing, CR-like positive neurons distributed in the lamina Ⅱ of the lumbar spinal cord increased slightly compared with the control group. After 3 days, there was no significant change in CR-like neurons compared with the control group. After 7 days, CR-like neurons decreased slightly. After 14 days, the expression of CR was significantly decreased. At 21 days, the expression of CR was recovered but still lower than that of 7 days. The gray value of CR immunoreactive products in the spinal cord posterior horn showed that the expression of CR was the lowest at 14 days after operation, which was statistically significant compared with the contralateral and control groups (P <0.05). Conclusion: The change of CR expression after sciatic nerve shows a certain spatial and temporal pattern, which provides experimental evidence for further revealing the role of CR in nervous system diseases.