香烟提出物对被动吸烟大鼠肺泡巨噬细胞源性明胶酶的表达和酶活性的影响  

Gelatinases expression and their activity in rat alveolar macrophages induced by cigarette smoke medium

作　　者：李亚清[1] 张珍祥[1] 徐永健[1] 陈仕新[1] 倪望[1] 杨朝[1] 马丹[1] 

机构地区：[1]华中科技大学同济医学院附属同济医院呼吸病研究室,湖北武汉430030

出　　处：《中国病理生理杂志》2006年第5期948-952,共5页Chinese Journal of Pathophysiology

摘　　要：目的:探讨香烟烟雾对被动吸烟大鼠肺泡巨噬细胞源性明胶酶A(MMP-2)和明胶酶B(MMP-9)表达及酶活性的影响。方法:复制被动吸烟大鼠模型,从其支气管肺泡灌洗液(BALF)中分离与培养大鼠肺泡巨噬细胞(AM),经Giemsa染色法鉴定,以台盼蓝染色计数和检测其活性;制备香烟提出物(CSM),刺激培养的AM。以半定量RT-PCR法检测MMP-2、MMP-9 mRNA表达水平;以明胶酶谱法检测MMP-2、MMP-9酶活性水平。结果:以0%、1%、3%、5%、10%、15%CSM刺激AM时,MMP-9和β-actin mRNA灰密度值比分别为:0.095±0.006、0.215±0.018、0.468±0.038、0.837±0.047、0.159±0.028、0.092±0.002;而MMP-2和β-actin mRNA灰密度值比分别为:0.109±0.023、0.176±0.036、0.317±0.024、0.370±0.051、0.274±0.025、0.087±0.005。MMP-2酶活性的相对灰密度值分别为6 007.83±147.39、38 202.00±3 160.40、70 532.00±3 225.18、90 074.00±925.62、70 070.17±2 060.33、17 487.67±2 060.33;MMP-9酶活性的相对灰密度值分别为10 904.67±1 697.005、2 627.83±3 974.05、95 284.83±3 098.891、03 400.50±3 441.309、4 650.33±3 825.08、27 571.17±2 129.48。组间均存在显著差异(P<0.05)。结论:香烟烟雾可诱导AM源性MMP-2和MMP-9 mRNA的表达,并导致其酶活性升高;AM及其分泌的明胶酶活性异常可能在吸烟相关的COPD中起重要作用。AIM： To study the effect of cigarette smoke medium （CSM） on the gene expression and activity of gelatinases from alveolar macrophages （AMs） in the rat, and then to explore their role in the pathogenesis of chronic obstructive pulmonary disease, （COPD）. METHODS： AMs were obtained from BALF of the rats that had smoked for 12 weeks. CSM was produced following the method of Wirtz and colleagues, and the cultured AMs were respectively stimulated for 24 h by 0%, 1%, 3%, 5%, 10%, 15% CSM. The mRNA levels of MMP - 9 and MMP-2 were detected by semi - quantitative RT- PCR, and the enzyme activity was measured by Zymography. RESULTS： When the concentration of CSM was below 5 %, the expression and activity of MMP - 9 and MMP - 2 signficanfly increased with the concentration of CSM in a dose - depended manner （ P 〈 0.05）. While the concentration of CSM exceeded 5 %, the expression and activity of MMP - 9 and MMP - 2 correspondingly decreased with the increase in CSM concentrations （P 〈 0.05）, which possibly were related with the cytotoxicity of CSM. CONCLUSION： The expression and activity of MMP - 9 and MMP - 2 are induced by CSM. The gelatinases induced by smoking from AM may play an important role in the pathogenesis of COPD.

关 键 词：巨噬细胞 肺泡 明胶酶类 烟雾 吸烟 

分 类 号：R363[医药卫生—病理学] R163[医药卫生—基础医学]