Objective To clarify the role of transforming growth factor‑β1 (TGF‑β1) in mechanical ventilation to promote aerobic glycolysis and accelerate the process of pulmonary fibrosis. Methods According to the random number table method, a total of 24 C57BL/6 mice were divided into the following groups (n=6): a sham operation group (Sham group), a vehicle control group (Vehicle group), a mechanical ventilation group (MV group) and a TGF‑β1 receptor inhibitor+mechanical ventilation (TGFβ‑Ri+MV) group. The Sham group only underwent intubation after anesthesia and maintained spontaneous breathing, while the Vehicle group was subjected to anesthesia intubation after intragastric administration of negative control vehicle for 5 days and maintained spontaneous breathing. The MV group underwent single mechanical ventilation with a tidal volume of 20 ml/kg and a ventilation frequency of 70 breaths per minute for 2 h. The TGFβ‑Ri+MV group was intragastrically administered with 10 mg/kg SB525334, a TGF‑β1 receptor inhibitor for 5 days, before single mechanical ventilation with a tidal volume of 20 ml/kg and a ventilation frequency of 70 breaths per minute for 2 h. Samples of each group were collected 7 days later. The content of TGF‑β1 in alveolar lavage fluid was detected by enzyme‑linked immunosorbent assay (ELISA). The amounts of lactate dehydrogenase A (LDHA) and collagen type Ⅰ α 1 chain (COL1A1) protein in lung tissue were detected by Western blot. The content of lactic acid in alveolar lavage fluid was detected by colorimetry. The degree of pulmonary fibrosis was observed by Masson staining.. Results Compared with the Sham group, the MV group presented increases in the content of TGF‑β1 (t=2.07, P=0.047), in the amounts of LDHA (t=5.09, P=0.003) and lactic acid (t=2.42, P=0.046), and in the content of COL1A1 protein (t=4.327, P=0.012), with increased collagen deposition by Masson staining. Compared with the MV group, the TGFβ‑Ri+MV group presented decreases in the amounts of LDHA (t=2.43, P=0.038) and lactic acid (t=3.16, P=0.025) and in the content of COL1A1 protein (t=5.78, P=0.004), with decreased collagen deposition by Masson staining. Conclusion Mechanical ventilation can induce the production of TGF‑β1 in lung tissue and promote aerobic glycolysis, which can accelerate the process of mechanical ventilation‑associated pulmonary fibrosis.