The objective of this study was to determine whether male and female bone-marrow-derived macrophages (BMDMs) exhibit distinct metabolic responses to inflammatory activation. We hypothesized that inflammatory stimulation would induce glycolytic reprogramming in BMDMs, but that the magnitude and direction of these metabolic shifts would differ between sexes. Bone marrow was flushed from the femur and tibia of adult C57BL/6J mice, and BMDMs were cultured for seven days with colony-stimulating factor-1 (50 ng/mL), passaged, and stimulated with lipopolysaccharide (LPS) plus interferon-gamma (IFN-γ) (100 ng/mL) for 24hr. The female cell count per plate was lower than the male cell count, which could point to different growth rates. Metabolic function was assessed using SeahorseTM extracellular flux analysis; global metabolite levels were measured by liquid chromatography mass spectrometry–based metabolomics; and transcriptomic changes were evaluated using bulk RNA sequencing. Both male and female BMDMs exhibited a significant increase in glycolysis and a decrease in mitochondrial oxidative phosphorylation following LPS + IFN-γ stimulation, as well as increased glycolytic proton efflux rate and reduced ATP-linked respiration in both sexes, with no significant sex differences in basal oxygen-consumption rate or maximal respiratory capacity. Metabolomics analysis revealed dysregulation of multiple glycolytic and pentose-phosphate pathway metabolites following LPS + IFN-γ stimulation. A principal component analysis (PCA) was performed, and it was found that the data clusters in the treatment groups for both males and females did not overlap, suggesting a strong effect of LPS + IFN-γ on the expression/level of the metabolites. Pair-wise PERMANOVA test showed significance (p = 0.005) between males and females within the treatment group, but not the controls. Metabolomics revealed elevated levels of glycolytic and pentose phosphate intermediates–including fructose-6-phosphate, dihydroxy-acetone-phosphate, hexose-phosphate, ribose-phosphate, L-arginine, succinate, glutamine, arginine, citrulline, D-sedoheptulose-1-7-phosphate, and fructose-1,6-bisphosphate–in both sexes, with minimal changes in tricarboxylic acid (TCA) cycle intermediates. Metabolomics also revealed a significant reduction of ethanolamine in both sexes. dAMP was reduced, but only in females. This was also observed in other metabolites, but those showed no significant differences within sexes, only between sexes. Sex differences were limited to fructose-1,6-bisphosphate (female: 23.63-fold; male: 8.28-fold; male vs. female: p = 0.002), ethanolamine (female: 0.41-fold; male: 0.74-fold; male vs. female: p = 0.02), and dAMP (female: 0.84-fold; male: 1.54-fold; male vs. female: p = 0.001). RNA sequencing revealed balanced up- and down-regulation of transcripts, accompanied by strong enrichment of inflammatory pathways and suppression of fatty acid oxidation genes. In summary, inflammatory activation induces a conserved metabolic reprogramming toward glycolysis and away from oxidative phosphorylation and fatty-acid metabolism in BMDMs, with only modest sex-dependent variation. These findings suggest that inflammatory metabolic remodeling is largely sex-independent at the cellular level. AHA Career Development Award 856365 Mouton and NIH/NHLBI R01 HL166737 Mouton This abstract was presented at the American Physiology Summit 2026 and is only available in HTML format. There is no downloadable file or PDF version. The Physiology editorial board was not involved in the peer review process.
Wright et al. (Fri,) studied this question.