Fig. 2

Blocking glycolysis alleviates lung injury in ALI mice by reducing M1 macrophage polarization and pyroptosis A-B: The expression levels of HK2, PKM2, LDHA in lung tissues were measured by Western blot. Immunoblot analysis of glycolytic protein HK2, PKM2, LDHA expression in lung, compared with the CON group. C: Detection of glycolysis-related genes Hk2, Pkm2, Ldha mRNA expression in the lung tissues by RT-qPCR. D-E: C57BL/6J mice were intraperitoneally injected with 2-DG 1 h before the LPS administration. Evaluation of the therapeutic effect of 2-DG on the lung tissue of ALI mice by HE staining (scale bar 100 μm). Inflammation score was measured. F: Lactate concentration in lung lysate was assayed. G:The ELISA assay was employed to quantify the concentration of IL-1β in the bronchoalveolar lavage fluid (BALF). H-I: Protein levels of NLRP3, GSDMD, IL-1β in lung were determined by Western blotting. J: Electron microscopy analyses membrane perforations of macrophages in lung tissue(scale bar 2.0 μm; scale bar 500 nm). K: IHC for iNOS in the lung tissue (scale bar 100 μm). The data shown are the means ± SEM obtained from three separate biological replicates. * P < 0.05, ** P < 0.01 by unpaired t test