Background and Purpose Chronic obstructive pulmonary diseases (COPD) are age-related, airflow-obstruction diseases mostly caused by cigarette smoke. However, the relationship between COPD and lung cellular senescence is still not fully understood. Here, we investigated how E3 ligase Pellino-1 mediated COPD and lung cellular senescence. Experimental Approach We used western blot, qPCR and co-IP assays to analyze the correlation of Pellino-1 and P21 in cells with or without silencing Pellino-1. Then we used flow cytometry, immunofluorescence staining and β- galactosidase assay to analyze the influences of silencing Pellino-1. Furthermore, we constructed COPD and aging models in vivo. Adenovirus of knock-down and overexpression Pellino-1 was used to infected mice. Immunohistochemistry and HE staining were used to analyze the lung pathology. Key Results Here, we first found that the E3 ubiquitin ligase Pellino-1 could bind to senescence marker p21 and modify p21 by K63-site ubiquitination and verified with silencing Pellino-1. Furthermore, we found that p21-mediated lung cellular senescence could be inhibited by silencing Pellino-1. Moreover, by constructing an adenovirus mouse model, we found that silencing Pellino-1 could inhibit COPD and inflammation via reduction of SASPs regulated by p21. Resistomycin, a potential Pellino-1 inhibitor, interrupts the interaction between Pellino-1 and p21, which accelerates the ubiquitin-dependent degradation of p21 and consequently inhibits lung cellular senescence and COPD progression. Conclusion and Implications Our study elucidated that inhibition of E3 ligase Pellino-1 exhibits therapeutic potential for treatment to attenuate the progression of lung cellular senescence and COPD.