Objective To study the protective effect of recombinant rat Club cell 16 (rCC16) protein on lunginjury and expressions of matrix metalloproteinase (MMP)-9 and tissue inhibitor of metalloproteinase (TIMP)-1 in micesuffering from chronic obstructive pulmonary disease (COPD). Methods C57BL/6 mice (n =40) were randomly dividedinto four groups: blank, COPD model, rCC16 treatment 1, and rCC16 treatment 2 groups. The mice were exposed totobacco smoke for 3 months to create a COPD model. The blank-group mice were exposed to room air. When the model wasstable, mice in the blank and model groups were administered phosphate-buffered saline (PBS; i.n.), and mice in theother two groups were treated with rCC16 (1. 0 or 2. 5 μg/ g body weight, i.n., respectively). The mental state, diet, bodyweightchanges and urine of mice were observed. Histological changes in the lung tissues in different groups were observedwith hematoxylin and eosin staining. mRNA and protein levels of MMP-9 and TIMP-1 were analyzed by quantitative reversetranscription-polymerase chain reaction and immunohistochemistry. Results Body weights in the COPD group weredecreased compared with those in the blank group, which were increased with the feeding period. Body weights in therCC16 treatment 2 group were increased obviously, whereas the body weights in the rCC16 treatment 1 group increased moreslowly, and the differences were significant ( P < 0. 05). Lung structure in the COPD group was changed with widenedinteralveolar septa and development of emphysema. rCC16 treatment alleviated those pulmonary alterations and reduced theformation of pulmonary bullae. The expressions of MMP-9 and TIMP-1 were significantly higher than those in the blankgroup ( P < 0. 05). rCC16 treatment inhibited the expressions of MMP-9 and TIMP-1, which were overexpressed in theCOPD group, and the differences were significant ( P < 0. 05). Moreover, the rCC16’s regulation on MMP-9 expressionwas dose-dependent. Conclusions Intranasal administration of rCC16 reduces the pulmonary injury and expressions ofMMP-9 and TIMP-1 in lung tissues of COPD mice. Our results demonstrate that rCC16 has a promising therapeutic effect against COPD.