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目的探讨NGF对哮喘小鼠肺及初级传入神经元(C7-T5脊神经节及相应脊髓后角)SH2-Bβ表达的调节作用。方法BALB/c小鼠30只,按随机数字表法分为正常对照组、哮喘组、anti-NGF组,每组10只。利用免疫组织化学方法,免疫印迹法(W estern B lot)测定各组SH2-Bβ的免疫反应变化;M etamoph图象分析系统对结果进行分析。结果免疫组织化学结果显示哮喘小鼠肺、C7-T5节段脊神经节及对应的脊髓后角SH2-Bβ免疫阳性产物平均光密度值(MOD)分别为0.806±0.023、0.766±0.018、0.547±0.014,明显高于对照组(0.243±0.018、0.131±0.011、0.215±0.029)(P<0.01)。而Anti-NGF组小鼠SH2-Bβ免疫阳性产物MOD分别为0.252±0.015、0.158±0.012、0.251±0.024,明显低于哮喘组(P<0.01)。W estern B lot显示哮喘小鼠的C7-T5节段脊髓及肺内SH2-BβMOD与-βactin MOD的比值(0.738±0.021和1.526±0.022)明显高于对照组(0.346±0.017和0.512±0.018,P<0.01);而Anti-NGF组小鼠在肺、C7-T5节段脊髓SH2-Bβ免疫阳性产物MOD与β-actioin MOD的比值(0.436±0.011和0.682±0.015)明显低于哮喘组(P<0.01)。结论肺内、C7-T5脊神经节及对应脊髓后角神经元的SH2-Bβ可能参与NGF介导的哮喘的发病过程。
Objective To investigate the regulatory effect of NGF on the expression of SH2-Bβ in lung and primary afferent neurons (C7-T5 spinal ganglia and corresponding spinal cord posterior horn) in asthmatic mice. Methods Thirty BALB / c mice were randomly divided into normal control group, asthma group and anti-NGF group according to random number table. Immunohistochemistry and Western Blot were used to detect the immune response of SH2-Bβ in each group. The results of M etamoph image analysis system were analyzed. Results Immunohistochemistry results showed that the mean optical density (MOD) values of SH2-Bβ immunoreactive products in the lung, C7-T5 segments of spinal cord and corresponding spinal dorsal horn of asthmatic mice were 0.806 ± 0.023,0.766 ± 0.018 and 0.547 ± 0.014 , Which was significantly higher than that of the control group (0.243 ± 0.018,0.131 ± 0.011,0.215 ± 0.029) (P <0.01). The SH2-Bβ immunoreactive products of anti-NGF group were 0.252 ± 0.015,0.158 ± 0.012,0.251 ± 0.024, which was significantly lower than that of asthma group (P <0.01). W estern B lot showed that the ratios of SH2-BβMOD to -βactin MOD in C7-T5 segment of asthmatic mice were significantly higher than those in control group (0.346 ± 0.017 and 0.512 ± 0.018, (0.436 ± 0.011 and 0.682 ± 0.015) in SH-Bβ immunoreactive products in the lung and C7-T5 segment of Anti-NGF group were significantly lower than those in the asthma group (P <0.01) P <0.01). Conclusions SH2-Bβ in the C7-T5 spinal ganglia and the corresponding spinal dorsal horn neurons may be involved in the pathogenesis of NGF-mediated asthma in lung.