哮喘豚鼠下呼吸道MMP-2的表达及NGF的调节作用  

Expression of MMP-2 and the Regulatory Effect of NGF in the Lower Respiratory Tract of the Asthmatic Guinea Pig

作　　者：刘晓湘[1] 曹德寿[1] 齐金萍[2] 方秀斌[1] 

机构地区：[1]中国医科大学基础医学院神经生物学教研室,辽宁沈阳110001 [2]沈阳医学院解剖学教研室,辽宁沈阳110034

出　　处：《解剖科学进展》2005年第4期307-310,i0002,共5页Progress of Anatomical Sciences

基　　金：国家自然科学基金资助项目(30070330);辽宁省教育厅基金资助项目(20121032)

摘　　要：目的探讨基质金属蛋白酶-2(MMP-2)在哮喘豚鼠下呼吸道的表达及神经生长因子(NGF)对MMP-2的调节作用。方法应用免疫组织化学方法检测各组豚鼠下呼吸道MMP-2免疫反应的变化。用W estern b lot蛋白印迹方法检测各组豚鼠肺组织NGF和MMP-2的表达。用Luzex-F实时图像分析系统和凝胶成像分析系统分别对结果进行图像分析。结果①免疫组织化学结果显示:豚鼠气道上皮和肺内炎性细胞的MMP-2阳性免疫反应产物的平均灰度值(0-深色,255-浅色),生理盐水组(185.60±4.81、161.47±5.71)与单纯致敏组(184.80±9.64、160.33±4.03)比较没有显著差异(P>0.05);哮喘组(141.93±4.66、129.77±4.07)明显低于生理盐水组和单纯致敏组(P<0.01);哮喘+NGF抗体组(鼻腔吸入NGF抗体,179.73±6.11、150.23±8.13)则明显高于哮喘组(P<0.01)。②W estern b lot结果显示:与生理盐水组和单纯致敏组比较,哮喘组豚鼠肺组织MMP-2和NGF阳性产物的IDV(Integrated Density Value)与β-actin IDV的比值均明显升高(P<0.01),而哮喘+NGF抗体组则明显低于哮喘组(P<0.01),表明NGF可上调MMP-2的表达。结论MMP-2可能是哮喘气道重塑的一种调控因素,NGF诱发MMP-2的表达可能是NGF参与哮喘发病的机制之一,NGF抗体抑制哮喘时下呼吸道MMP-2的表达有可能延缓气道重塑的发生,可能是治疗哮喘的新途径。Objective To explore the expression of matrix metaUoproteinase-2（MMP-2） and the regulatory effect of nerve growth factor （NGF） in the lower respiratory tract of the asthmatic guinea pig. Methods The changes of MMP-2 immunoreactivity were investigated by means of immunohistocbemistry in lower respiratory tract of the guinea pigs in different groups. The expressions of NGF and MMP-2 were observed by Western blot in the lung of all the groups. The results were analyzed by the Luzex-F real time image analysis system and the gel imaging analysis system respectively. Results ①Immunohistochemistry results showed that there was no obvious difference for the mean grey values （0 - black to 255-white） of MMP-2 positive products in the airway epithelia and the pulmonary inflammatory cells between normal saline control group（ 185.60± 4.81,161.47± 5.71 ） and simply sensitized group （ 184.80 ± 9.64,160.33 ± 4.03） （ P 〉 0.05 ）. The mean grey values of MMP-2 positive products decreased significandy in the lower respiratory tract （141.93 ±4.66,129.77 ± 4.07 ）of the asthmatic guinea pigs compared with the normal saline control group and the simply sensitized group （ P 〈0.01 ）. However, the mean grey values were much higher in NGF antibody group（ inhalation of NGF antibody into nasal cavity, 179.73 ± 6.11,150.23 ± 8.13） titan in asthmatic group （ P 〈0.01 ）. ②Western blot results showed that： compared with normal saline control group and simply sensitized group, the ratios between Integrated Density Value （IDV） for MMP-2 or NGF protein band and 1DV for β-actin protein band increased markedly in tire lung of asthmatic group （ P 〈0. 01 ） , however, the ratios of NGF antibody group reduced markedly in comparison witb asthmatic group （ P 〈0.01 ） , indicating that NGF can upregulate the expression of MMP-2. Conclusions MMP -2 might play a role in airway remodeling in asthma, the overex- pression of MMP-2 induced by NGF might be one of tire mechanisms for the invo

关 键 词：基质金属蛋白酶-2 神经生长因子 下呼吸道 哮喘 豚鼠 

分 类 号：R562.25[医药卫生—呼吸系统]