Establishment of rat model for aspiration pneumonia and potential mechanisms

Abstract Background Aspiration pneumonia is a severe health concern, particularly for ICU patients with impaired airway defenses. Current animal models fail to fully replicate the condition, focusing solely on chemical lung injury from gastric acid while neglecting pathogen‐induced inflammation. This gap hinders research on pathogenesis and treatment, creating an urgent need for a clinically relevant model. This study aimed to develop an improved rat model of aspiration pneumonia by combining hydrochloric acid (HCl) and lipopolysaccharide (LPS) administration. Methods Specific pathogen‐free Sprague Dawley rats underwent intratracheal instillation of HCl and LPS. Techniques included rat weight measurement, tracheal intubation, pulmonary function monitoring, lung tissue sampling with HE staining and scoring, bronchoalveolar lavage fluid (BALF) sampling, protein and inflammatory cytokine analysis via BCA and ELISA, BALF pH determination, Evans Blue dye assessment, blood gas analysis, FITC‐dextran leakage, Western blotting, electron microscopy, survival analysis, and transcriptome sequencing with bioinformatics. Statistical analysis was performed using GraphPad Prism. Results The optimal model involved instillation of 1.5 μL/g.wt HCl (pH = 1) followed by 20 μg/g.wt LPS after 1 h. This model reproduced acute lung injury, including tissue damage, pulmonary microvascular dysfunction, inflammatory responses, hypoxemia, and impaired pulmonary ventilation, with recovery observed at 72 h. PANoptosis was confirmed, characterized by increased markers. Concentration‐dependent effects of HCl and LPS on lung damage were identified, alongside cytokine elevation and microvascular dysfunction. Conclusions This optimized model closely mimics clinical aspiration pneumonia, providing a valuable tool for studying pathophysiology and therapeutic strategies.