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)...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。展开更多
BACKGROUND： Several studies have demonstrated that SH2-B13 is over-expressed in the lung, C7T5 spinal ganglia, and the corresponding spinal dorsal horn of asthmatic mice. SH2-Bβ expression has been shown to positive...BACKGROUND： Several studies have demonstrated that SH2-B13 is over-expressed in the lung, C7T5 spinal ganglia, and the corresponding spinal dorsal horn of asthmatic mice. SH2-Bβ expression has been shown to positively correlate with nerve growth factor （NGF） expression levels. This indicates that SH2-Bβ, in the presence of NGF, may participate in asthmatic attacks. OBJECTIVE： To observe the effects of anti-NGF on SH2-Bβ expression in primary afferent neurons （C7-T5 spinal ganglia and corresponding spinal dorsal horn） and in the lung to further investigate the regulatory effects of NGF on SH2-B/3 expression. DESIGN, TIME AND SETTING： A completely randomized block design experiment was performed at the Department of Neurobiology, China Medical University between March 2004 and July 2005. MATERIALS： Thirty-six male, BALB/c mice were included in this study. Ovalbumin solution was purchased from Sigma, USA. SH2-Bβ polyclonal antibody was provided by Santa Cruz, USA. Anti-NGF reagent was obtained from Wuhan Boster Bioengineering Co., Ltd., China. METHODS： Thirty-six mice were randomly and evenly divided into three groups： control, model, and anti-NGF. In the model group, asthma was induced by intraperitoneal injection and aerosol inhalation of ovalbumin solution. Mice in the anti-NGF group received anti-NGF through the nasal cavity 3 hours prior to aerosol inhalation. In the control group, mice were subjected to experimental procedures similar to the model group, except that ovalbumin solution was replaced by phosphate buffered saline （PBS）. MAIN OUTCOME MEASURES： SH2-Bβ expression in primary afferent neurons （C7T5 spinal ganglia and the corresponding spinal dorsal horn） and the lung was detected by immunohistochemistry and Western blot. Immunostaining intensity level of SH2-Bβ was analyzed using the MetaMorph image analysis system. RESULTS： lmmunohistochemistry results revealed that the mean intensity of SH2-Bβ expression in the C7 T5 spinal ganglia, the corresponding spinal dorsal horn, and the lungs was significantly greater in the model group than in the control group （P 〈 0.01）. However, expression was significantly less in the anti-NGF group, compared with the model group （P 〈 0.01）. Western Blot results demonstrated that SH2-Bβ expression was significantly greater in the model group C7-T5 spinal ganglia and lung, compared with the control group （P 〈 0.01）. However, expression was significantly less in the anti-NGF group, compared with the model group （P 〈 0.01）. CONCLUSION： The present study showed that, in the primary afferent neurons and the lung, SH2-Bβ participated in asthmatic attack. Anti-NGF down-regulated SH2-Bβ expression in C7-T5 spinal ganglia and the corresponding spinal dorsal horn, as well as the lung, of asthmatic mice. These results indicate that SH2-Bβ could be an important signaling molecule in mediating effects of NGF during asthmatic attack.展开更多
A previous study by our research group showed that nerve growth factor is involved in the onset of asthma through regulating SH2-Bβ expression in the lung and visceral primary afferent neurons of asthmatic mice. This...A previous study by our research group showed that nerve growth factor is involved in the onset of asthma through regulating SH2-Bβ expression in the lung and visceral primary afferent neurons of asthmatic mice. This study sought to assess the expression level of interleukin-1β in primary afferent neurons in C7-T5 spinal ganglia, spinal cord and lung in asthmatic mice after blockage of SH2-Bβ. The levels of interleukin-1β protein in primary afferent neurons in the C7-T5 spinal ganglia and lung were decreased, and interleukin-1β mRNA expression also down-regulated in the spinal cord, medulla oblongata and lung tissue after blockage of SH2-Bβ. Our findings indicate that SH2-Bβ can upregulate the expression of interleukin-1β in C7-T5 spinal ganglia, spinal cord and lung of asthmatic mice.展开更多
文摘目的探讨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。
文摘BACKGROUND： Several studies have demonstrated that SH2-B13 is over-expressed in the lung, C7T5 spinal ganglia, and the corresponding spinal dorsal horn of asthmatic mice. SH2-Bβ expression has been shown to positively correlate with nerve growth factor （NGF） expression levels. This indicates that SH2-Bβ, in the presence of NGF, may participate in asthmatic attacks. OBJECTIVE： To observe the effects of anti-NGF on SH2-Bβ expression in primary afferent neurons （C7-T5 spinal ganglia and corresponding spinal dorsal horn） and in the lung to further investigate the regulatory effects of NGF on SH2-B/3 expression. DESIGN, TIME AND SETTING： A completely randomized block design experiment was performed at the Department of Neurobiology, China Medical University between March 2004 and July 2005. MATERIALS： Thirty-six male, BALB/c mice were included in this study. Ovalbumin solution was purchased from Sigma, USA. SH2-Bβ polyclonal antibody was provided by Santa Cruz, USA. Anti-NGF reagent was obtained from Wuhan Boster Bioengineering Co., Ltd., China. METHODS： Thirty-six mice were randomly and evenly divided into three groups： control, model, and anti-NGF. In the model group, asthma was induced by intraperitoneal injection and aerosol inhalation of ovalbumin solution. Mice in the anti-NGF group received anti-NGF through the nasal cavity 3 hours prior to aerosol inhalation. In the control group, mice were subjected to experimental procedures similar to the model group, except that ovalbumin solution was replaced by phosphate buffered saline （PBS）. MAIN OUTCOME MEASURES： SH2-Bβ expression in primary afferent neurons （C7T5 spinal ganglia and the corresponding spinal dorsal horn） and the lung was detected by immunohistochemistry and Western blot. Immunostaining intensity level of SH2-Bβ was analyzed using the MetaMorph image analysis system. RESULTS： lmmunohistochemistry results revealed that the mean intensity of SH2-Bβ expression in the C7 T5 spinal ganglia, the corresponding spinal dorsal horn, and the lungs was significantly greater in the model group than in the control group （P 〈 0.01）. However, expression was significantly less in the anti-NGF group, compared with the model group （P 〈 0.01）. Western Blot results demonstrated that SH2-Bβ expression was significantly greater in the model group C7-T5 spinal ganglia and lung, compared with the control group （P 〈 0.01）. However, expression was significantly less in the anti-NGF group, compared with the model group （P 〈 0.01）. CONCLUSION： The present study showed that, in the primary afferent neurons and the lung, SH2-Bβ participated in asthmatic attack. Anti-NGF down-regulated SH2-Bβ expression in C7-T5 spinal ganglia and the corresponding spinal dorsal horn, as well as the lung, of asthmatic mice. These results indicate that SH2-Bβ could be an important signaling molecule in mediating effects of NGF during asthmatic attack.
基金grant from the Liaoning Provincial Education Bureau, No. 20060890
文摘A previous study by our research group showed that nerve growth factor is involved in the onset of asthma through regulating SH2-Bβ expression in the lung and visceral primary afferent neurons of asthmatic mice. This study sought to assess the expression level of interleukin-1β in primary afferent neurons in C7-T5 spinal ganglia, spinal cord and lung in asthmatic mice after blockage of SH2-Bβ. The levels of interleukin-1β protein in primary afferent neurons in the C7-T5 spinal ganglia and lung were decreased, and interleukin-1β mRNA expression also down-regulated in the spinal cord, medulla oblongata and lung tissue after blockage of SH2-Bβ. Our findings indicate that SH2-Bβ can upregulate the expression of interleukin-1β in C7-T5 spinal ganglia, spinal cord and lung of asthmatic mice.
