国际呼吸杂志
國際呼吸雜誌
국제호흡잡지
INTERNATIONAL JOURNAL OF RESPIRATION
2009年
1期
9-12,封3
,共5页
王巧玲%陈金龙%齐金萍%方秀斌
王巧玲%陳金龍%齊金萍%方秀斌
왕교령%진금룡%제금평%방수빈
哮喘%神经生长因子%磷脂酶C%小鼠%气道阻力
哮喘%神經生長因子%燐脂酶C%小鼠%氣道阻力
효천%신경생장인자%린지매C%소서%기도조력
Asthma%Nerve growth factor%Phospholipase C%Mice%Airway resistance
目的 研究支气管哮喘(简称哮喘)小鼠下呼吸道及内脏感觉传入部位磷脂酶C-γ(PLC-γ)的表达,及神经生长因子(nerve growth factor,NGF)对PLC-γ在上述部位的调节作用,探讨NGF介导的信号转导通路在哮喘发病机制中的作用.方法 BABL/C 30只,按随机数字表法分为正常对照组、哮喘组及NGF阻断组.利用AniRes2005肺功能仪测BABL/C小鼠气道阻力,应用免疫组织化学方法和免疫印记法(Western blotting),观察PLC-γ在哮喘小鼠下呼吸道及内脏感觉传人部位的表达及NGF被阻断后PLC-γ表达水平的变化,Metamoph图象分析系统对结果进行分析.结果 气道阻力检测结果显示,哮喘组小鼠较正常对照组气道阻力明显增高(P<0.01),NGF阻断组气道阻力均较哮喘组明显降低(P<0.01).免疫组织化学结果显示PLC在NGF阻断组小鼠肺组织(0.273±0.018),C7～T5段脊神经节(0.158±0.012),以及相应节段的脊髓后角内(0.168±0.022)表达明显低于哮喘组(0.423±0.023,0.351±0.018,0.368±0.014)(P<0.01).Western blotting结果显示NGF阻断组肺(0.921±0.014)及C7～T5节段脊髓(0.835±0.023)PLC-γ MOD值与β-actin MOD值的比值明显低于哮喘组(1.476±0.017,1.251±0.019)(P<0.01).结论 PLC-γ在哮喘小鼠肺、C7～T5节段脊神经节及相应的脊髓后角过表达,NGF上调哮喘上述部位PLC-γ的表达,提示NGF可通过调节肺内及内脏传人部位PLC的表达参与哮喘发病过程.
目的 研究支氣管哮喘(簡稱哮喘)小鼠下呼吸道及內髒感覺傳入部位燐脂酶C-γ(PLC-γ)的錶達,及神經生長因子(nerve growth factor,NGF)對PLC-γ在上述部位的調節作用,探討NGF介導的信號轉導通路在哮喘髮病機製中的作用.方法 BABL/C 30隻,按隨機數字錶法分為正常對照組、哮喘組及NGF阻斷組.利用AniRes2005肺功能儀測BABL/C小鼠氣道阻力,應用免疫組織化學方法和免疫印記法(Western blotting),觀察PLC-γ在哮喘小鼠下呼吸道及內髒感覺傳人部位的錶達及NGF被阻斷後PLC-γ錶達水平的變化,Metamoph圖象分析繫統對結果進行分析.結果 氣道阻力檢測結果顯示,哮喘組小鼠較正常對照組氣道阻力明顯增高(P<0.01),NGF阻斷組氣道阻力均較哮喘組明顯降低(P<0.01).免疫組織化學結果顯示PLC在NGF阻斷組小鼠肺組織(0.273±0.018),C7～T5段脊神經節(0.158±0.012),以及相應節段的脊髓後角內(0.168±0.022)錶達明顯低于哮喘組(0.423±0.023,0.351±0.018,0.368±0.014)(P<0.01).Western blotting結果顯示NGF阻斷組肺(0.921±0.014)及C7～T5節段脊髓(0.835±0.023)PLC-γ MOD值與β-actin MOD值的比值明顯低于哮喘組(1.476±0.017,1.251±0.019)(P<0.01).結論 PLC-γ在哮喘小鼠肺、C7～T5節段脊神經節及相應的脊髓後角過錶達,NGF上調哮喘上述部位PLC-γ的錶達,提示NGF可通過調節肺內及內髒傳人部位PLC的錶達參與哮喘髮病過程.
목적 연구지기관효천(간칭효천)소서하호흡도급내장감각전입부위린지매C-γ(PLC-γ)적표체,급신경생장인자(nerve growth factor,NGF)대PLC-γ재상술부위적조절작용,탐토NGF개도적신호전도통로재효천발병궤제중적작용.방법 BABL/C 30지,안수궤수자표법분위정상대조조、효천조급NGF조단조.이용AniRes2005폐공능의측BABL/C소서기도조력,응용면역조직화학방법화면역인기법(Western blotting),관찰PLC-γ재효천소서하호흡도급내장감각전인부위적표체급NGF피조단후PLC-γ표체수평적변화,Metamoph도상분석계통대결과진행분석.결과 기도조력검측결과현시,효천조소서교정상대조조기도조력명현증고(P<0.01),NGF조단조기도조력균교효천조명현강저(P<0.01).면역조직화학결과현시PLC재NGF조단조소서폐조직(0.273±0.018),C7～T5단척신경절(0.158±0.012),이급상응절단적척수후각내(0.168±0.022)표체명현저우효천조(0.423±0.023,0.351±0.018,0.368±0.014)(P<0.01).Western blotting결과현시NGF조단조폐(0.921±0.014)급C7～T5절단척수(0.835±0.023)PLC-γ MOD치여β-actin MOD치적비치명현저우효천조(1.476±0.017,1.251±0.019)(P<0.01).결론 PLC-γ재효천소서폐、C7～T5절단척신경절급상응적척수후각과표체,NGF상조효천상술부위PLC-γ적표체,제시NGF가통과조절폐내급내장전인부위PLC적표체삼여효천발병과정.
Objective To study the expression of phospholipase C-γ(PLC-γ) in lower respiratory tract and visceral sensation afferent site of asthmatic mice and the regulatory effect of nerve growth factor(NGF) on PLC-γ in above-mentioned site,to explore the function of NGF mediated signal pathway in the asthmatic pathogenesis. Methods Thirty BALB/c mice were randomly divided into normal control group, asthmatic group,anti-NGF group on average. The airway resistance of mice was measured by AniRes 2005 lung function meter. By means of immunohistochemistry and Western blotting to investigate the changes of PLC-γ immunoreactivity and the regulatory effect of NGF in the lower respiratory tract and viscerosensory afferent sites of the asthmatic mice model and analyzed by Metamoph anaysis system. Results Airway resistance measurement showed: increased airway resistance to methacholine was observed in the asthmatic group compared with the normal control group, but decreased in anti-NGF group compared with the asthmatic group(P<0.01). The expression of PLC-γ was decreased significantly in lower respiratory tract (0. 273 ± 0.018) and visceral sensory afferent sites[C7-T5 spinal ganglia (0. 158±0. 012) and the corresponding posterior horn of the spinal cord (0. 168±0. 022)] of the anti-NGF group compared with the asthmatic mice model (0. 423±0. 023,0. 351±0.018,0. 368±0. 014) (P<0. 01). The result of Western blotting shows that the ratio between PLC-γ MOD value of lung (0. 921± 0. 014 ), C7-T5 spinal cord (0. 835 ± 0. 023 ) and β-actin MOD value in anti-NGF group is obviously decreased to that of the asthmatic group(1. 476±0. 017, 1. 251±0. 019). Conclusions There is overexpression of PLC-γ in the lungs, C7-T5 spinal ganglia and corresponding spinal dorsal horn. NGF upregulates the expression of PLC-γ in the above-mentioned site. This finding suggests that PLC-γ might take part in the mechanism of NGF mediated asthma.
