Fig. 5: TLR4 ablation promotes metabolic reprogramming and mitochondrial homeostasis in septic macrophages.

A The AUC scores for glycolysis, OXPHOS, and positive ROS from scRNA-seq analysis across the four groups. Increased glycolysis and ROS production were observed in the WT CLP group, while the Tlr4^-/- CLP group showed higher OXPHOS activity compared to WT CLP. B The relative lactate levels in macrophages from the BALF of each mouse group (n  =  5). C Flow cytometry quantifying glucose uptake (FITC-2-NBDG) across the four groups (n  =  5). D The NADH levels in macrophages from the BALF of each mouse group (n  =  5). E Bar graph illustrating the NAD⁺ / NADH ratio, indicating a shift towards glycolysis in the WT CLP group (n  =  5). F Relative amounts of intracellular ATP in WT and Tlr4^-/- macrophages (n  =  5). G Levels of ROS were measured in macrophages from the BALF via flow cytometry (n  =  5). H JC-1 staining of mitochondrial membrane potential in macrophages from each group. The WT CLP group shows greater mitochondrial dysfunction (green staining) compared to the other groups, with Tlr4^-/- CLP showing more preserved membrane potential. Data are represented as mean ± SD, ***P < 0.001.