神经生长因子对哮喘豚鼠内脏感觉传入部位MMP-2表达的影响  

The Upregulative Effect of NGF on MMP-2 in the Viscerosensory Afferent Sites 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》2006年第1期38-41,i0005,共5页Progress of Anatomical Sciences

基　　金：国家自然科学基金资助项目(30570811)

摘　　要：目的 探讨基质金属蛋白酶-2（MMP-2）在哮喘豚鼠内脏感觉传入部位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豚鼠内脏感觉传入部位MMP-2免疫反应的变化。用Western blot蛋白印迹方法检测豚鼠C7～T5段脊神经节和相应节段脊髓背角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C7～T5段脊神经节和相应节段脊髓背角内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（175．50±4．67、166．47±6．20）与单纯致敏组（172．63±9．51、165．20±5．14）比较没有显著差异（P〉0．05）；哮喘组（139．87±4．88、125．93±4．49）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，169．73±6．24、152．37±8．67）则明显高于哮喘组（P〈0．01）。②Western blot结果显示：与生理盐水组和单纯致敏组比较，哮喘组豚鼠C7-T5段脊神经节及相应节段脊髓样品中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可上调豚鼠内脏感觉传入部位MMP-2的表达。结论 C 7～T5段脊神经节以及相应节段脊髓背角内的MMP-2可能也参与哮喘的发病过程，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MMP-2的表达有可能廷缓气道重塑的发生，可能是治疗哮喘的新途径。Objective To explore the expression of matrix metalloproteinase-2（MMP-2） and the regulatory effect of nerve growth factor （NGF） in the viscerosensory afferent sites of the asthmatic guinea pig. Methods The changes of MMP-2 immtmore.aetivity were investigated by means of immunohistochemistry in the viscerosensory afferent sites of the guinea pigs in different groups. The expression of NGF and MMP-2 were observed by Western blot in the C7 - T5 spinal ganglia and the corresponding spinal dorsal horn 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 （1）Immunohistochemistry 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 C7 - T5 spinal ganglia and the corresponding spinal dorsal horn between normal saline control group（175.50 ±4.67, 166.47±6.20）and simply sensitized group（172.63 ±9.51, 165.20 ±5.14） （ P 〉 0.05）. The mean grey values of MMP-2 positive products decreased slgnificantly in the C7 -T5 spinal ganglia（139. 87 ±4.88）and the corresponding spinal dorsal horn（ 125.93 ±4.49）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, 169.73 ± 6.24,152.37 ± 8.67 ） than in asthmatic group （ P 〈0.01 ）.（2）Western 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 IDV for β-actin protein band increased markedly in the C7 - T5 spinal ganglia and the corresponding spinal dorsal horn of asthmatic group （ P 〈0.01）. However, the ratios of IDV for NGF antibody group reduced markedly in comparison with asthmatic group （ P 〈 0.01 ）, indicating that NGF can upregula

关 键 词：基质金属蛋白酶-2 神经生长因子 脊神经节 脊髓背角 哮喘 豚鼠 

分 类 号：R562.25[医药卫生—呼吸系统] R322.8[医药卫生—内科学]