Objective To construct an acute lung injury mouse model induced by cholic acid with intratracheal instillation or intranasal infusion, and to screen the perfusion method and verify the feasibility of bile acid-induced lung injury. Methods Mice were grouped according to the factorial design of administration method × drug ( 3 × 3). The administration method included tracheotomy, nasal drops for 1 day, and nasal drops for 6 days. Animals were administered either cholic acid, DMSO, or PBS; in total, there were nine groups of mice. During the administration period, body weight changes were monitored. After administration, chest X-ray examination was performed, histological observation of the pathological changes of the lung tissue was carried out, arterial blood was taken to analyze the partial pressure of oxygen (PO₂), and ELISA was used to detect tumor necrosis in the lung tissue, as demonstrated by tumor necrosis factor-α (TNF-α) and interleukin 1β ( IL-1β) expression. Results The X-ray result of the intratracheal instillation and reperfusion cholic acid group showed diffuse infiltration of lung tissue. Gross lung samples showed obvious hemorrhage, while histopathology showed a large amount of inflammatory cell infiltration and alveolar wall thickening. Blood oxygen partial pressure was decreased, and TNF-α and IL-1β were significantly higher than in the other groups. The indexes of the cholic acid group after 1 day of intranasal infusion and 6 days of intranasal infusion were not as good as those in the cholic acid group after intratracheal instillation and reperfusion. Conclusions The acute lung injury model can be successfully constructed by intratracheal instillation of cholic acid.