Chronic obstructive pulmonary disease (COPD) development is the result of environmental factors interact with hereditary factors, and smoking is the primary cause for COPD development. Nevertheless, both the mechanisms of smoking leads to COPD and the mechanisms of COPD hereditary susceptibility are not well clarified so far. Proteomics research features high efficiency and rich information, which had provided strong help for COPD study, and considered has broad prospects in COPD research area. Two dimensional gel electrophoresis and matrix assisted laser desorption/ionization time of flight mass spectrometry were used in proteomics research to compare the lung tissue proteome of never-smokers, non-COPD smokers and COPD smokers. By combined with bioinformatics technology, 24 proteins were identified to be differentially expressioned between groups. The D4-GDI expression level in lung tissue of non-COPD smokers was 1.7 times to never-smokers, while the D4-GDI expression level in lung tissue of COPD smokers was nearly twice to non-COPD smokers. For verification, D4-GDI expression level in lung tissue was detected by immunohistochemical staining and Western-blotting, and the results was consistent with proteomics research. The results of this study for the first time to description: Smoking can up-regulate D4-GDI expression level in lung tissue, D4-GDI involved in the pathogenesis of COPD and may be associated with COPD susceptibility.