Background: Acute lung injury (ALI) and its severe manifestation, acute respiratory distress syndrome, are life-threatening pulmonary disorders for which effective therapeutic interventions are currently lacking. This study aimed to investigate the role of ferroptosis in LPS-induced ALI and elucidate the synergistic mechanism of metallothionein-1 (MT-1) and galactoxylan polysaccharide (VDPS) in inhibiting ferroptosis, potentially offering novel therapeutic strategies for ALI.

Methods: Using Lipopolysaccharide (LPS)-induced murine models and MLE-12 cell lines, we analyzed ferroptosis-related indicators, including glutathione peroxidase 4 (GPX4), malondialdehyde (MDA), reduced glutathione (GSH), and iron content. Interventions with ferroptosis inhibitor ferrostatin-1 (Fer-1), activator erastin, and VDPS were performed. MT-1 knockdown experiments were conducted to explore its role in oxidative stress, iron metabolism, and inflammatory responses.

Results: LPS stimulation induced characteristic ferroptosis features in both mice and MLE-12 cells, manifested by increased pulmonary iron content, decreased GSH levels, elevated MDA levels, and reduced GPX4 expression (p < 0.05). Fer-1 treatment significantly ameliorated these oxidative stress markers and protected lung function (p < 0.05). LPS markedly upregulated MT-1 expression, while MT-1 knockdown exacerbated oxidative stress, lipid peroxidation, and ferroptosis (p < 0.05). VDPS effectively inhibited LPS-induced ferroptosis, reducing oxidative damage and inflammation (p < 0.05). Combined treatment with Fer-1 notably decreased pulmonary edema, inflammatory infiltration, and fibrosis (p < 0.05). VDPS also regulated iron metabolism, suppressing elevated iron levels in lung tissue and serum (p < 0.05). Importantly, MT-1 enhanced VDPS-mediated GPX4 pathway activation, synergistically inhibiting ferroptosis and attenuating LPS-induced oxidative injury (p < 0.05).

Conclusion: This study reveals the crucial role of ferroptosis in LPS-induced ALI. MT-1 enhances VDPS-mediated ferroptosis inhibition by modulating oxidative stress responses and antioxidant enzyme activities, effectively alleviating lung injury. These findings provide experimental evidence for MT-1 and VDPS as potential therapeutic strategies, highlighting the promising application of anti-ferroptosis therapy in ALI treatment.