Abstract
Hyperuricemia (HUA) is a metabolic disorder characterized by elevated blood uric acid (UA) levels, closely associated with conditions such as gout. UA-induced macrophage M1 polarization fundamentally exacerbates inflammatory pathophysiology, but current HUA-specific immunoregulatory treatments are inadequate. This study investigates whether β-hydroxybutyrate (BHB), a UA-degrading metabolite produced by Lacticaseibacillus rhamnosus M2b, can suppress UA-induced M1 macrophage polarization and promote M2 polarization via activation of the GPR109a–AMPK signaling axis, aiming to identify novel targets for intervening in HUA-related inflammation. Using untargeted metabolomics combined with CCK-8 and ELISA assays, BHB was identified as the key active molecule. An M1 polarization model was established by stimulating RAW264.7 cells with 1 mM UA. Experimental groups included control, UA, UA + BHB, and UA + M2b-conditioned medium (UA + M2b_CM) groups. Results showed that both BHB and M2b_CM significantly inhibited UA-induced M1 polarization, as indicated by reduced levels of IL-1β, IL-6, TNF-α, and iNOS. Concurrently, they promoted M2 polarization markers CD163 and IL-10 expression, and enhanced AMPK phosphorylation (increased p-AMPK/AMPK ratio) (P < 0.05). It was confirmed by transcriptomic analysis that BHB intervention could significantly enhance the activity of the AMPK signaling pathway. Functional validation experiments demonstrated that either silencing GPR109a expression with small interfering RNA (siRNA) or inhibiting AMPK with Compound C completely abolished the anti-inflammatory effects of BHB. Conversely, overexpression of GPR109a enhanced the anti-inflammatory efficacy of BHB. This result confirms that GPR109a is the primary molecular target mediating the effects of BHB, and that GPR109a participates in this mediating process through the AMPK signaling pathway. In conclusion, BHB derived from Lacticaseibacillus rhamnosus M2b inhibits UA-induced M1 macrophage polarization and promotes M2 polarization by activating the GPR109a–AMPK signaling pathway. These findings provide a new strategic perspective for utilizing gut microbiota metabolites in the treatment of HUA-related inflammation.
Similar content being viewed by others
Data availability
The metabolomics datasets generated and analysed during the current study are available in the MetaboLights repository under accession code MTBLS13224. All data can be publicly accessed via the following persistent link: [https://www.ebi.ac.uk/metabolights/MTBLS13224]. All original experimental data generated for this work are available in the Supplementary Materials of this article.
Abbreviations
- HUA:
-
Hyperuricemia
- UA:
-
uric acid
- BHB:
-
β-hydroxybutyrate
- M2b_CM:
-
M2b Conditioned Medium
- GPR109a:
-
G protein - coupled receptors 109a
- AMPK:
-
AMP-activated protein kinase
- HMTA:
-
2-Hydroxy-3-methylbutyric acid
- 3-HP:
-
3-Hydroxypropionic acid
- 3-4-HPPA:
-
3-(4-Hydroxyphenyl)propionic acid
- 3-HPAA:
-
3-Hydroxyphenylacetic acid
- N-acetyl GABA:
-
4-Acetamidobutyric acid
References
Li, L., Zhang, Y. & Zeng, C. Update on the epidemiology, genetics, and therapeutic options of hyperuricemia. Am. J. Transl Res. 12, 3167–3181 (2020).
Hu, H. et al. The SGLT2 inhibitor Dapagliflozin ameliorates renal fibrosis in hyperuricemic nephropathy. Cell. Rep. Med. 5, 101690. https://doi.org/10.1016/j.xcrm.2024.101690 (2024).
Crișan, T. O. et al. Soluble uric acid primes TLR-induced Proinflammatory cytokine production by human primary cells via Inhibition of IL-1Ra. Ann. Rheum. Dis. 75, 755–762. https://doi.org/10.1136/annrheumdis-2014-206564 (2016).
Xu, W. et al. The protective effects of neoastilbin on monosodium urate stimulated THP-1-Derived macrophages and gouty arthritis in mice through NF-κB and NLRP3 inflammasome pathways. Molecules 27 https://doi.org/10.3390/molecules27113477 (2022).
Aikepa, D. et al. The effects of specific gut microbiota on Hyperuricemia - A Mendelian randomization analysis and clinical validation. Diabetes Metab. Syndr. Obes. 18, 1891–1902. https://doi.org/10.2147/dmso.S510384 (2025).
Chen, W. et al. Safety evaluation of Human-Derived uric acid degrading Lacticaseibacillus paracasei M2a and its impact on gut microbiota. Probiotics Antimicrob. Proteins. https://doi.org/10.1007/s12602-025-10562-x (2025).
Yamada, N. et al. Lactobacillus gasseri PA-3 uses the purines IMP, inosine and hypoxanthine and reduces their absorption in rats. Microorganisms 5 https://doi.org/10.3390/microorganisms5010010 (2017).
Park, H. K. & Lee, S. J. Treatment of gouty arthritis is associated with restoring the gut microbiota and promoting the production of short-chain fatty acids. Arthritis Res. Ther. 24 https://doi.org/10.1186/s13075-022-02742-9 (2022).
Martínez-Nava, G. A. et al. The impact of short-chain fatty acid-producing bacteria of the gut microbiota in hyperuricemia and gout diagnosis. Clin. Rheumatol. 42, 203–214. https://doi.org/10.1007/s10067-022-06392-9 (2023).
Kayama, H., Okumura, R. & Takeda, K. Interaction between the Microbiota, Epithelia, and immune cells in the intestine. Annu. Rev. Immunol. 38, 23–48. https://doi.org/10.1146/annurev-immunol-070119-115104 (2020).
Liu, H. et al. Oil mistparticulate matter exposure induces hyperlipidemia-related inflammation via microbiota/ SCFAs/GPR43 axis inhibition and TLR4/NF-κB activation. Environmental pollution (Barking, Essex:) 344, 123331, ) 344, 123331, (1987). https://doi.org/10.1016/j.envpol.2024.123331 (2024).
Qi, J. et al. Low glucose plus β-Hydroxybutyrate induces an enhanced inflammatory response in Yak alveolar macrophages via activating the GPR109A/NF-κB signaling pathway. Int. J. Mol. Sci. 24 https://doi.org/10.3390/ijms241411331 (2023).
Huang, Y. et al. [GPR109A partly mediates inhibitory effects of β-hydroxybutyric acid on lung adenocarcinoma cell proliferation, migration and invasion]. Nan Fang Yi Ke Da Xue Xue Bao. 43, 1744–1751. https://doi.org/10.12122/j.issn.1673-4254.2023.10.12 (2023).
Thio, C. L., Lai, A. C., Ting, Y. T., Chi, P. Y. & Chang, Y. J. The ketone body β-hydroxybutyrate mitigates ILC2-driven airway inflammation by regulating mast cell function. Cell. Rep. 40, 111437. https://doi.org/10.1016/j.celrep.2022.111437 (2022).
Deng, G., Wen, B., Jia, L., Liu, J. & Yan, Q. Clostridium Butyricum upregulates GPR109A/AMPK/PGC-1α and ameliorates acute pancreatitis-associated intestinal barrier injury in mice. Arch. Microbiol. 206, 265. https://doi.org/10.1007/s00203-024-04001-8 (2024).
Guo, W. et al. GPR109A alleviate mastitis and enhances the blood milk barrier by activating AMPK/Nrf2 and autophagy. Int. J. Biol. Sci. 17, 4271–4284. https://doi.org/10.7150/ijbs.62380 (2021).
Youm, Y. H. et al. The ketone metabolite β-hydroxybutyrate blocks NLRP3 inflammasome-mediated inflammatory disease. Nat. Med. 21, 263–269. https://doi.org/10.1038/nm.3804 (2015).
Bradshaw, P. C., Seeds, W. A., Miller, A. C., Mahajan, V. R. & Curtis, W. M. COVID-19: proposing a Ketone-Based metabolic therapy as a treatment to blunt the cytokine storm. Oxid. Med. Cell. Longev. 2020 (6401341). https://doi.org/10.1155/2020/6401341 (2020).
Aikepa, D. et al. Isolation, identification, whole genome sequencing, and functional analysis of a highly efficient human-derived uric acid-degrading bacterium, M2a. Microbiol. Bull. 1–16. https://doi.org/10.13344/j.microbiol.china.240460 (2024). (in China).
Terkeltaub, R. & Dodd, D. The gut Microbiome in hyperuricemia and gout. Arthritis Rheumatol. 77, 955–965. https://doi.org/10.1002/art.43118 (2025).
Song, S. et al. Alteration of gut Microbiome and correlated amino acid metabolism contribute to hyperuricemia and Th17-Driven inflammation in Uox-KO mice. Front. Immunol. 13, 804306. https://doi.org/10.3389/fimmu.2022.804306 (2022).
Xu, Y. X. et al. Alistipes indistinctus-derived Hippuric acid promotes intestinal urate excretion to alleviate hyperuricemia. Cell. Host Microbe. 32, 366–381e369. https://doi.org/10.1016/j.chom.2024.02.001 (2024).
Quaresma, M. et al. Probiotic mixture containing Lactobacillus spp. And bifidobacterium spp. Attenuates 5-fluorouracil-induced intestinal mucositis in mice. Nutr. Cancer. 72, 1355–1365. https://doi.org/10.1080/01635581.2019.1675719 (2020).
Abdelrahman, A. A. et al. Expression and activation of the ketone body receptor HCAR2/GPR109A promotes preservation of retinal endothelial cell barrier function. Exp. Eye Res. 221, 109129. https://doi.org/10.1016/j.exer.2022.109129 (2022).
Sun, Y. et al. Parabacteroides distasonis ameliorates insulin resistance via activation of intestinal GPR109a. Nat. Commun. 14, 7740. https://doi.org/10.1038/s41467-023-43622-3 (2023).
Qiu, X., Wu, W., Zhang, S., Huang, C. & Lin, D. 3-Hydroxybutyrate promotes myoblast proliferation and differentiation through energy metabolism and GPR109a-Mediated Ca(2+)-NFAT signaling pathways. J. Proteome Res. 24, 2063–2080. https://doi.org/10.1021/acs.jproteome.4c01150 (2025).
Zou, H. et al. Lipid catabolism in starved Yak is inhibited by intravenous infusion of β-Hydroxybutyrate. Anim. (Basel). 10 https://doi.org/10.3390/ani10010136 (2020).
Lei, J., Shu, Z., Zhu, H. & Zhao, L. A. M. P. K. Regulates M1 macrophage polarization through the JAK2/STAT3 signaling pathway to attenuate airway inflammation in Obesity-Related asthma. Inflammation 48, 372–392. https://doi.org/10.1007/s10753-024-02070-x (2025).
Li, C., Zhang, H., Liu, Y., Zhang, T. & Gu, F. Gpr109A in TAMs promoted hepatocellular carcinoma via increasing PKA/PPARγ/MerTK/IL-10/TGFβ induced M2c polarization. Sci. Rep. 15, 18820. https://doi.org/10.1038/s41598-025-02447-4 (2025).
Lee, A. K., Kim, D. H., Bang, E., Choi, Y. J. & Chung, H. Y. β-Hydroxybutyrate suppresses lipid accumulation in aged liver through GPR109A-mediated signaling. Aging Dis. 11, 777–790. https://doi.org/10.14336/ad.2019.0926 (2020).
Chen, Y. et al. β-Hydroxybutyrate alleviates atherosclerotic calcification by inhibiting Endoplasmic reticulum Stress-Mediated apoptosis via AMPK/Nrf2 pathway. Nutrients 17 https://doi.org/10.3390/nu17010111 (2024).
Jayashankar, S. S., Tajul Arifin, K. & Nasaruddin, M. L. β-Hydroxybutyrate regulates activated microglia to alleviate neurodegenerative processes in neurological diseases: A scoping review. Nutrients 15 https://doi.org/10.3390/nu15030524 (2023).
Zhai, X. et al. AMPK-regulated glycerol excretion maintains metabolic crosstalk between reductive and energetic stress. Nat. Cell. Biol. 27, 141–153. https://doi.org/10.1038/s41556-024-01549-x (2025).
Funding
This study gratefully acknowledges the following institutions: the National Natural Science Foundation of China (Project Numbers: 82260182 and 82360180); the Xinjiang Uygur Autonomous Region Natural Science Foundation (Project Numbers: 2024D01C129) for their critical financial support, and the authors thank the State Key Laboratory of Pathogenesis, Prevention, and Treatment of High-Incidence Diseases in Central Asia and the Laboratory of Molecular Biology of Endemic Diseases, Xinjiang Medical University, for providing all necessary equipment.
Author information
Authors and Affiliations
Contributions
M.D. and Y.H. contributed equally to this work. M.D.: conceptualization, data curation, and writing. Y.H.: methodology and data curation. Y.Z. and X.C. were responsible for methodology and investigation; J.S. and B.Z. contributed to methodology; D.Y. performed validation; Q.Y. managed data curation; W.C. and Y.S. oversaw the project as supervisors and acquired funding. All authors have reviewed and approved the final manuscript.
Corresponding author
Ethics declarations
Competing interests
The authors declare no competing interests.
Additional information
Publisher’s note
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Supplementary Information
Below is the link to the electronic supplementary material.
Rights and permissions
Open Access This article is licensed under a Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License, which permits any non-commercial use, sharing, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if you modified the licensed material. You do not have permission under this licence to share adapted material derived from this article or parts of it. The images or other third party material in this article are included in the article’s Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by-nc-nd/4.0/.
About this article
Cite this article
Du, M., He, Y., Zhu, Y. et al. GPR109a-AMPK axis mediates the Attenuation of uric acid-induced M1 macrophage polarization by β-hydroxybutyrate from Lacticaseibacillus rhamnosus M2b. Sci Rep (2026). https://doi.org/10.1038/s41598-026-39746-3
Received:
Accepted:
Published:
DOI: https://doi.org/10.1038/s41598-026-39746-3
