UNLABELLED: Macrophages are critical components of the innate immune system that adapt their function based on local microenvironmental cues. In adipose tissue, macrophage polarization is intimately linked to metabolic regulation and inflammation. In this pilot study, we performed lipidomic profiling to characterize the phospholipid composition of pro-inflammatory (PI-ATM) and non-inflammatory (NI-ATM) macrophages isolated from visceral adipose tissue of living kidney donors. Tissue samples were processed, and macrophages were isolated and sorted using CD14 and CD16 markers. Lipids were extracted by methyl tert-butyl ether, and differences in the lipidome were analyzed using the optimized UHPLC-MS method. PI-ATM exhibited a higher abundance of lipids across various lipid classes than NI-ATM. Phosphatidylethanolamines and ether lipids derived from phosphatidylethanolamines and phosphatidylcholines were most frequently found among the significantly altered lipids. The majority of significantly altered lipid species contained an octadecenoic aliphatic chain. These findings suggest that alterations in membrane lipid composition - particularly ether lipids and phosphatidylethanolamines - may play a pivotal role in macrophage polarization and adipose tissue inflammation. Our study provides novel insights into the lipidomic regulation of macrophage function in human adipose tissue and identifies potential targets for therapeutic intervention in metabolic disorders. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1038/s41598-025-32912-z.

Department of Analytical Chemistry Faculty of Science Charles University Prague Prague Czech Republic

Department of Analytical Chemistry University of Chemistry and Technology Prague Prague Czech Republic

Department of Anesthesiology Resuscitation and Intensive Care Medicine 1st Faculty of Medicine Charles University and Military University Hospital Prague Czech Republic

Department of Epidemiology and Biostatistics 3 Faculty of Medicine Charles University Prague Czech Republic

Department of Physiology Faculty of Science Charles University Prague Prague Czech Republic

Institute for Clinical and Experimental Medicine Prague Czech Republic

Institute of Organic Chemistry and Biochemistry Czech Academy of Sciences Prague Czech Republic

Zobrazit více v PubMed

Lazarov, T., Juarez-Carreño, S., Cox, N. & Geissmann, F. Physiology and diseases of tissue-resident macrophages. PubMed PMC

Azzu, V., Vacca, M., Virtue, S., Allison, M. & Vidal-Puig, A. Adipose tissue-liver cross talk in the control of whole-body metabolism: Implications in nonalcoholic fatty liver disease. PubMed

Kosteli, A. et al. Weight loss and lipolysis promote a dynamic immune response in murine adipose tissue. PubMed PMC

Jaitin, D. A. et al. Lipid-Associated macrophages control metabolic homeostasis in a Trem2-Dependent manner. PubMed PMC

Poledne, R., Kralova Lesna, I., Kralova, A., Fronek, J. & Cejkova, S. The relationship between non-HDL cholesterol and macrophage phenotypes in human adipose tissue. PubMed PMC

Kralova Lesna, I. et al. Cardiovascular disease predictors and adipose tissue macrophage polarization: is there a link? PubMed

Lesna, K. I. et al. Macrophage subsets in the adipose tissue could be modified by sex and the reproductive age of women. PubMed

Poledne, R. et al. Polarization of macrophages in human adipose tissue is related to the fatty acid spectrum in membrane phospholipids. PubMed PMC

Bohdanowicz, M. & Grinstein, S. Role of phospholipids in endocytosis, phagocytosis, and macropinocytosis. PubMed

Fessler, M. B. & Parks, J. S. Intracellular lipid flux and membrane microdomains as organizing principles in inflammatory cell signaling. PubMed PMC

Yeudall, S. et al. Macrophage acetyl-CoA carboxylase regulates acute inflammation through control of glucose and lipid metabolism. PubMed PMC

Morgan, P. K. et al. Macrophage polarization state affects lipid composition and the channeling of exogenous fatty acids into endogenous lipid pools. PubMed PMC

Fang, H. et al. Lipopolysaccharide-induced macrophage inflammatory response is regulated by SHIP. PubMed

Cauvi, D. M., Hawisher, D., Derunes, J., Rodriguez, E. & De Maio, A. Membrane phospholipids activate the inflammatory response in macrophages by various mechanisms. PubMed

Bartuskova, H. et al. Links between macrophages in perivascular adipose tissue and arterial wall: a role in atherosclerosis initiation? PubMed

Matyash, V., Liebisch, G., Kurzchalia, T. V., Shevchenko, A. & Schwudke, D. Lipid extraction by methyl-tert-butyl ether for high-throughput lipidomics. PubMed PMC

Tsugawa, H. et al. MS-DIAL: data-independent MS/MS Deconvolution for comprehensive metabolome analysis. PubMed PMC

Cajka, T. et al. Exploring the impact of organic solvent quality and unusual adduct formation during LC-MS-Based lipidomic profiling. PubMed PMC

Heymsfield, S. B. & Wadden, T. A. Mechanisms, Pathophysiology, and management of obesity. PubMed

Rakusanova, S. & Cajka, T. Tips and tricks for LC-MS-based metabolomics and lipidomics analysis.

Wallner, S. et al. Phosphatidylcholine and phosphatidylethanolamine plasmalogens in lipid loaded human macrophages. PubMed PMC

Kralova Lesna, I. et al. Characterisation and comparison of adipose tissue macrophages from human subcutaneous, visceral and perivascular adipose tissue. PubMed PMC

Cífková, R. et al. 30-year trends in major cardiovascular risk factors in the Czech population, Czech MONICA and Czech post-MONICA, 1985–2016/17. PubMed PMC

Nagan, N. & Zoeller, R. A. Plasmalogens: biosynthesis and functions. PubMed

Bozelli, J. C. Jr., Azher, S. & Epand, R. M. Plasmalogens and chronic inflammatory diseases. PubMed PMC

Jiménez-Rojo, N. & Riezman, H. On the road to unraveling the molecular functions of ether lipids. PubMed

Glaser, P. E. & Gross, R. W. Plasmenylethanolamine facilitates rapid membrane fusion: a stopped-flow kinetic investigation correlating the propensity of a major plasma membrane constituent to adopt an HII phase with its ability to promote membrane fusion. PubMed

Banerjee, S. et al. The Vps13-like protein BLTP2 is pro-survival and regulates phosphatidylethanolamine levels in the plasma membrane to maintain its fluidity and function. PubMed PMC

Wallner, S. et al. Monocyte to macrophage differentiation goes along with modulation of the plasmalogen pattern through transcriptional regulation. PubMed PMC

Hill, D. A. et al. Distinct macrophage populations direct inflammatory versus physiological changes in adipose tissue. PubMed PMC

Papin, M. et al. Endogenous ether lipids differentially promote tumor aggressiveness by regulating the SK3 channel. PubMed PMC

Benjamin, D. I. et al. Ether lipid generating enzyme AGPS alters the balance of structural and signaling lipids to fuel cancer pathogenicity. PubMed PMC

Schooneveldt, Y. L., Paul, S., Calkin, A. C. & Meikle, P. J. Ether lipids in obesity: from cells to population studies. PubMed PMC

Jové, M. et al. Ether lipid-mediated antioxidant defense in alzheimer’s disease. PubMed PMC

Cedillo, L. et al. Ether lipid biosynthesis promotes lifespan extension and enables diverse pro-longevity paradigms in PubMed PMC

Hornburg, D. et al. Dynamic lipidome alterations associated with human health, disease and ageing. PubMed PMC

Valtersson, C., Filipsson, L. & Dallner, G. Localization of phosphatidylethanolamine in microsomal membranes and regulation of its distribution by the fatty acid composition. PubMed

Prades, J., Funari, S. S., Escribá, P. V. & Barceló, F. Effects of unsaturated fatty acids and triacylglycerols on phosphatidylethanolamine membrane structure. PubMed

Santos, M. et al. The non-enzymatic oxidation of phosphatidylethanolamine and phosphatidylserine and their intriguing roles in inflammation dynamics and diseases. PubMed

Li, Z. et al. The ratio of phosphatidylcholine to phosphatidylethanolamine influences membrane integrity and steatohepatitis. PubMed

Petkevicius, K. et al. TLCD1 and TLCD2 regulate cellular phosphatidylethanolamine composition and promote the progression of non-alcoholic steatohepatitis. PubMed PMC

Santa-María, C. et al. Update on anti-inflammatory molecular mechanisms induced by oleic acid. PubMed PMC

Santamarina, A. B. et al. Anti-inflammatory effects of oleic acid and the anthocyanin Keracyanin alone and in combination: effects on monocyte and macrophage responses and the NF-κB pathway. PubMed

Kim, D. H., Cho, Y. M., Lee, K. H., Jeong, S. W. & Kwon, O. J. Oleate protects macrophages from palmitate-induced apoptosis through the downregulation of CD36 expression. PubMed

Suganami, T., Nishida, J. & Ogawa, Y. A paracrine loop between adipocytes and macrophages aggravates inflammatory changes: role of free fatty acids and tumor necrosis factor alpha. PubMed

Rubio, J. M., Astudillo, A. M., Casas, J., Balboa, M. A. & Balsinde, J. Regulation of phagocytosis in macrophages by membrane ethanolamine plasmalogens. PubMed PMC

Gil-de-Gómez, L., Astudillo, A. M., Lebrero, P., Balboa, M. A. & Balsinde, J. Essential role for ethanolamine plasmalogen hydrolysis in bacterial lipopolysaccharide priming of macrophages for enhanced arachidonic acid release. PubMed PMC

Arroyo, A. B. et al. Peds1 deficiency in zebrafish results in myeloid cell apoptosis and exacerbated inflammation. PubMed PMC