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目的 揭示喉返神经损伤及修复后疑核运动神经元睫状神经营养因子 (Ciliaryneurotrophicfactor,CNTF)的表达及分布。方法 实验犬分为喉返神经损伤组和损伤修复组 ,采用原位杂交及免疫组织化学技术 ,结合图像分析技术测定疑核CNTFmRNA及其蛋白表达呈阳性反应的神经元胞体灰度、体积及数量。结果 疑核运动神经元自分泌CNTF ,神经损伤后早期胞体内CNTF基因表达下降而CNTF蛋白表达增强。 4周后基因表达迅速增强 ,而CNTF蛋白表达减弱 ,但 6周后基因及其蛋白表达和神经元胞体数及体积均始终低于健侧。 12周后CNTF表达、神经元形态、胞体数及体积趋向稳定。神经修复早期 ,CNTF表达与神经损伤相似 ,但基因表达恢复较神经损伤组快 ,CNTF蛋白高表达维持时间较神经损伤组长 ,尽管 12周后表达的神经元较正常减少 ,但表达强度及体积与正常无明显差异。结论 运动神经元表达CNTF ;表达程度与神经损伤及再生有关 ,提示CNTF在神经损伤再生中起重要作用。
Objective To reveal the expression and distribution of ciliary neurotrophic factor (CNTF) after injury and repair of recurrent laryngeal nerve. Methods The experimental dogs were divided into the recurrent laryngeal nerve injury group and the injury repair group. The in situ hybridization and immunohistochemical techniques were used to determine the gray scale, volume and number of neuronal cell bodies with suspected nuclear CNTF mRNA and protein expression by image analysis . Results The autocrine CNTF of suspected motoneurons decreased the expression of CNTF gene in early period of post-nerve injury and the expression of CNTF increased. After 4 weeks, the gene expression increased rapidly and the expression of CNTF protein decreased. However, the expression of the gene and its protein and the number and volume of neurons were always lower than that of the contralateral side after 6 weeks. After 12 weeks, CNTF expression, neuronal morphology, cell number and volume tend to be stable. At the early stage of nerve repair, the expression of CNTF was similar to that of nerve injury, but the expression of CNTF was faster than that of nerve injury group. The expression of CNTF protein was longer than that of nerve injury group. Although the number of neurons decreased after 12 weeks, the expression intensity and volume No significant difference with normal. Conclusions The expression of CNTF in motor neurons is related to nerve injury and regeneration, suggesting that CNTF plays an important role in the regeneration of nerve injury.