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目的研究SH2-Bβ在哮喘小鼠肺及内脏感觉传入系统(C7~T5脊神经节及对应的脊髓后角)的表达,探讨SH2-Bβ在哮喘发病中的作用。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,每组15只。利用免疫组织化学方法测定两组小鼠肺、C7~T5脊神经节及对应的脊髓后角SH2-Bβ的免疫反应变化;用免疫印迹法(Western blotting)检测肺及C7~T5节段脊髓SH2-Bβ及神经生长因子(NGF)的免疫反应变化;Metamoph图像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)。Western blotting显示SH2-Bβ及NGF在哮喘小鼠的C7~T5节段脊髓及肺内表达明显强于对照组;正常对照组脊髓及肺内的平均相对光密度值分别为0.346±0.017和0.512±0.018,哮喘组为0.738±0.021和1.526±0.022。NGF在正常对照组脊髓及肺内的平均相对光密度值分别为0.357±0.021和0.734±0.013,哮喘组为1.445±0.015和1.265±0.018;且SH2-Bβ与NGF光密度值呈正相关(r=0.651,P<0.01)。结论结果提示,SH2-Bβ在哮喘小鼠肺、C7~T5节段脊神经节及对应的脊髓后角过表达,可能是NGF参与哮喘发病的重要信号分子。
Objective To investigate the expression of SH2-Bβ in lung and visceral afferent system (C7 ~ T5 spinal ganglia and corresponding spinal dorsal horn) of asthmatic mice, and to explore the role of SH2-Bβ in the pathogenesis of asthma. Methods Thirty BALB / c mice were randomly divided into normal control group and asthma group according to random number table. Immunohistochemical staining was used to detect the immunoreactivity of SH2-Bβ in the lung, C7 ~ T5 spinal ganglia and corresponding spinal dorsal horn in both groups. Western blotting was used to detect the expression of SH2- Bβ and nerve growth factor (NGF) immune response changes; Metamoph image analysis system to analyze the results. Results The results of immunohistochemistry showed that the average optical density of SH2-Bβ immunoreactive products in the lung, C7 ~ T5 segments of spinal cord and the corresponding spinal cord posterior horn of asthmatic mice were 0.806 ± 0.023,0.766 ± 0.018 and 0.547 ± 0.014, respectively. (0.243 ± 0.018,0.131 ± 0.011,0.215 ± 0.029) in the control group (P <0.01). The results of Western blotting showed that the expression of SH2-Bβ and NGF in spinal cord and lung of C7 ~ T5 in asthmatic mice was significantly stronger than that in control group. The average relative optical density in spinal cord and lung of normal control group were 0.346 ± 0.017 and 0.512 ± 0.018 for the asthma group and 0.738 ± 0.021 for the asthma group and 1.526 ± 0.022 for the asthma group. The mean relative optical density of NGF in the normal control group was 0.357 ± 0.021 and 0.734 ± 0.013 in the spinal cord and lung respectively, and 1.445 ± 0.015 and 1.265 ± 0.018 in the asthma group. The SH2-Bβ was positively correlated with the optical density of NGF (r = 0.651, P <0.01). Conclusions The results suggest that over-expression of SH2-Bβ in the lung, C7 ~ T5 segmental spinal ganglia and corresponding spinal dorsal horn of asthmatic mice may be an important signaling molecule involved in the pathogenesis of asthma.