全反式维甲酸与弹性蛋白酶对肺组织破坏的干预作用  

Inhibiting Effects of Retinoic Acid and Elastase on Lung Tissue Destruction

作　　者：王舒[1] 朱运奎[1] 闫圣杰[2] 李继东[1] 金远林[1] 刘杜娇[1] 汤育瑛[1] 

机构地区：[1]兰州军区兰州总医院呼吸内科,甘肃省兰州市730050 [2]兰州大学第二临床医学院

出　　处：《中国全科医学》2010年第8期827-830,共4页Chinese General Practice

基　　金：甘肃省自然科学基金项目暨中青年科技项目(3ZS051-A25-088)

摘　　要：目的采用胚肺成纤维细胞三维立体培养技术,直接观察全反式维甲酸(RA)和中性粒细胞弹性蛋白酶(NE)对肺的主要成分之一胶原组织降解的干预作用。方法在胚肺成纤维细胞三维立体培养过程中加入细胞因子〔肿瘤坏死因子-α(TNF-α)和白介素-1β(IL-1β)〕来诱导基质金属蛋白酶(MMPs)的表达,同时加入RA或NE干预MMPs及其组织抑制因子(TIMPs)的表达;用明胶酶谱法测定MMP-2、MMP-9,用Western blotting法测定MMP-1、MMP-3及TIMP-1、TIMP-2,同时在培养5d后测定胶原的含量及面积大小。结果TNF-α和IL-1β诱导在立体胶原内培养的胚肺成纤维细胞产生MMP-1、MMP-3和MMP-9,但是只引起少量胶原降解(P>0.05),同时加入NE,结果导致胶原几乎完全降解(P<0.01)。NE使MMP-1、MMP-2、MMP-3和MMP-9由非活化的形式转化为分子量较小的活化形式,同时,清除TIMP-1和TIMP-2。RA抑制了MMP-1的表达及MMP-3、MMP-9的活化,并削弱了NE对TIMP-1、TIMP-2的清除作用,进而抑制了胶原的降解。胶原降解后基质发生强烈收缩。结论MMPs可直接降解肺内胶原和其他间质组织,导致肺组织破坏;NE通过活化MMPs引起或加速细胞外基质的破坏;MMPs与NE的协同作用可能是肺气肿等肺组织破坏的机制。RA通过抑制MMPs的表达与活化对抗NE对肺组织的破坏作用。Objective To investigate the effects of all-trans retinoic acid （ATRA） and neutrophil elastase （NE） on lung fibroblast mediated collagen degradation using embryonic lung fibroblast 3-dimensional culture technique.Methods Human lung fibroblasts were cultured in three dimensional collagen gels,then TNF-αand IL-1βwere added to induce expression of matrix metalloproteinases （MMPs）,and ATRA or NE added at the same time to the culture media.The expressions of MMP-2,MM-9 were detected by gelatin zymography,and MMP-1,MMP-3 and MMP （TIMP）-1,TIMP-2 by Western blotting.Collagen content and gel areas were determined after 5 days of culture.Results TNF-αand IL-1β induced lung fibroblast cultured in 3-dimensional collagen to produce MMP-1,3,9,but caused only a little collagen degradation （P〉0.05）.However,adding NE at the same time led to complete collagen degradation （P〈0.01）.NE activated MMP-1,MMP-2,MMP-3 and MMP-9 into active forms,and cleared TIMP-1 and TIMP-2.ATRA inhibited the expression of MMP-1 and the activation of MMP-3 and MMP-9,and reduced the elimination of TIMP-1 and TIMP-2 by NE.The intense matrix contraction appeared after collagen degradation.Conclusion MMPs may directly degrade the pulmonary collagen and other interstitial tissues to result in the lung tissue destruction,NE can induce or accelerate the destruction of extracellular matrices by activating MMPs.The synergistic interaction between NE and MMPs may be involved in lung tissue destruction of emphysema.ATRA may inhibit the expression and activation of MMPs thus to reduce the lung tissue destruction by NE.

关 键 词：全反式维甲酸 白细胞弹性蛋白酶 基质金属蛋白酶 成纤维细胞 胶原 三维立体培养 

分 类 号：R329.453[医药卫生—人体解剖和组织胚胎学]