Objective Lumbar spinal needle, indwelling needle, and tracheotomy were used to establish ableomycin-induced pulmonary fibrosis model in C57BL/6J mice,respectively,and to compare the effects of these threemethods on mice. Methods One hundred and twenty C57BL/6J mice were randomly divided into blank, lumbar spinalneedle model, indwelling needle model, and tracheotomy model groups. Mice from different groups were administered byintratracheal instillation of bleomycin using the different methods, respectively. The general condition of the mice (changesin body weight and survival rate) was observed, and the histopathological changes (HE staining and Masson staining) andhydroxyproline level were examined. Results The body weight of the lumbar spinal needle model group and the indwellingneedle model group was significantly higher than that of the tracheotomy model group at 28 days after modeling ( P <0. 01? P <0. 001). There were no significant differences in survival rate among the four groups except between the blank group andthe tracheotomy model group ( P <0. 05). The degrees of alveolitis and pulmonary fibrosis were highest in the tracheotomymodel group, followed by the lumbar spinal needle model group and indwelling needle model group. The degree of alveolitisand pulmonary fibrosis was greatest at 21 days after modeling. The lesions were evenly distributed in the tracheotomy modelgroup, and unevenly distributed in the lumbar spinal needle model and indwelling needle model groups. The lesions in thelung tissue around the trachea were more severe than in other areas in the lumbar spinal needle model and indwelling needlemodel groups. The level of hydroxyproline in the lumbar spinal needle model and tracheotomy model groups was significantlyhigher than that in the blank group ( P <0. 01, P <0. 001). Conclusions In summary, tracheal intubation by lumbar spinal needle is the best method among the three methods for the establishment of a bleomycin-induced pulmonary fibrosis in mice.