Daucosterol Targets PFKFB3 to Mitigates Sepsis-Induced Acute Lung Injury by Inhibiting Glycolysis and M1 Macrophage Polarization.

Abstract

Acute lung injury (ALI) is a severe inflammatory condition often triggered by infections. Glycolysis and macrophage polarization play critical roles in ALI pathogenesis. This study investigates the effects of Daucosterol on lipopolysaccharide (LPS)-stimulated lung injury and its potential mechanisms. In this research, BEAS-2B lung epithelial cells and MH-S alveolar macrophages were exposed to LPS and various concentrations of Daucosterol. Cell viability was assessed using CCK-8 assay. Glycolytic activity was evaluated by detecting ATP production, lactate level, and extracellular acidification rate. Macrophage polarization was assessed using flow cytometry. Tumor necrosis factor (TNF)-α and interleukin (IL)-6 levels were detected by enzyme-linked immunosorbent assay. Gene expression was evaluated by reverse transcription-quantitative polymerase chain reaction and western blotting. A cecal ligation and puncture (CLP)-induced ALI mouse model was used to validate the protective effect of Daucosterol in vivo. Results showed that Daucosterol prevented the reduction in cell viability in LPS-stimulated BEAS-2B cells. In MH-S macrophages, Daucosterol inhibited LPS-induced glycolysis and M1 polarization while promoting M2 polarization. Mechanistically, Daucosterol reversed the LPS-induced upregulation of PFKFB3. Overexpression of PFKFB3 counteracted the inhibitory effects of Daucosterol on glycolysis and M1 polarization. In vivo, Daucosterol significantly alleviated CLP-induced ALI by improving lung histopathology, reducing pulmonary edema, enhancing oxygenation, inhibiting myeloperoxidase and caspase-3 activity, and decreasing TNF-α and IL-6 levels in bronchoalveolar lavage fluid. In conclusion, Daucosterol targets PFKFB3 to mitigate sepsis-induced ALI by inhibiting glycolysis and M1 macrophage polarization, offering a potential therapeutic strategy for ALI.