SH2-Bβ抗体对哮喘小鼠肺及初级传入神经元SH2-Bβ表达的调节作用

The Regulatory Effect of Anti-SH2-Bβ on the Expression of SH2Bβ in the Lung and Primary Afferent Neurons of the Asthmatic Mice

目的:探讨SH2-Bβ在哮喘发病中的作用。方法:用卵蛋白致敏致喘的方法制备BALB/c小鼠哮喘模型。应用免疫组织化学方法和免疫印记法(Western blot)观察SH2-Bβ在哮喘小鼠下呼吸道及内脏感觉传入部位的表达及应用SH2-Bβ抗体后SH2-Bβ水平的变化,Metamoph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)。而应用SH2-Bβ抗体组相应部位的平均光密度值分别为(0.252±0.015、0.158±0.012、0.251±0.024)明显低于哮喘组(P<0.01)。Western blot结果显示:哮喘组肺及C7-T5节段脊髓后角SH2-Bβ与β-actin MOD值的比(2.512±0.021,2.615±0.022)明显高于对照组(0.675±0.019,0.712±0.018)(P<0.01),而应用SH2-Bβ抗体组相应部位的比值(0.969±0.023,1.023±0.022)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参与哮喘的发病过程,anti-SH2-Bβ可下调SH2-Bβ的表达,减轻肺部炎症。

Objective： To study the effect of SH2-Bβ on asthma. Methods： Asthma was induced in BALB/c mice by exposure to chicken egg ovalbumin （OVA）. Immunohistochemistry and Western blot were used to investigate the changes of SH2-Bβ immunoreactivity and the regulatory effect of anti- SH2-Bβ in the lower respiratory tract and viscerosensory afferent sites of the asthmatic mice. And results were analyzed by the Metamoph analysis system. Results： Immunohistochemistry demonstrated that the mean optic density （MOD） of SH2-Bβ positive substance in the lungs, C7-T5 spinal ganglia and the corresponding posterior horn of the spinal cord （0. 806 ± 0. 023, 0. 766 ± 0. 018, 0. 547 ± 0. 014） of the asthmatic mice was much higher than those （ 0. 243 ± 0. 018, 0. 131 ± 0. 011, 0. 215 ±0. 029） in normal control group respectively （ P 〈 0. 01 ）, but MOD values （0. 252 ± 0. 015, 0. 158 ± 0. 012, 0. 251±0. 024 ） respectively in anti- SH2-Bβ group were much lower than those in the asthmatic mice （P 〈0. 01 ）. Western Blot demonstrated that the mean ratio of SH2-Bβ MOD to β-actin MOD in lung and spinal cord was （2. 512 ±0. 021, 2.615 ±0.022） in asthmatic group, much higher than （0. 675 ± 0. 019, 0. 712 ± 0. 018 ） in normal control group （ P 〈 0. 01 ） ; and those （0. 969 ± 0. 023, 1. 023 ±0. 022） in anti- SH2-Bβ group was much lower than those of asthmatic mice （P〈0. 01 ）. Conclusion： The present results indicate that the SH2-Bβ in the lungs, primary afferent neurons （ C7-T5 spinal ganglia and the corresponding posterior horn of the spinal cord） might be involved in the pathogenesis of asthma. Anti- SH2-Bβ down-regulates the expression of the SH2-Bβ in these portion and reduce the inflammation in the lungs of asthmatic mice.