Unraveling anti-inflammatory metabolic signatures of Glycyrrhiza uralensis and isoliquiritigenin through multiomics

1. Che, C. T., Wang, Z. J., Chow, M. S. & Lam, C. W. Herb-herb combination for therapeutic enhancement and advancement: theory, practice and future perspectives. Molecules 18, 5125–5141 (2013).

   Google Scholar 

2. Zhou, X. et al. Synergistic effects of Chinese herbal medicine: a comprehensive review of methodology and current research. Front. Pharm. 7, 201 (2016).

   Google Scholar 

3. Adamski, Z., Blythe, L. L., Milella, L. & Bufo, S. A. Biological activities of alkaloids: from toxicology to pharmacology. Toxins 12 https://doi.org/10.3390/toxins12040210 (2020).

4. Roy, A. et al. Flavonoids a bioactive compound from medicinal plants and its therapeutic applications. Biomed. Res. Int. 2022, 5445291 (2022).

   Google Scholar 

5. Rao, A. V. & Gurfinkel, D. M. The bioactivity of saponins: triterpenoid and steroidal glycosides. Drug Metab. Drug Interact. 17, 211–235 (2000).

   Google Scholar 

6. Honda, H. et al. Glycyrrhizin and isoliquiritigenin suppress the LPS sensor toll-like receptor 4/MD-2 complex signaling in a different manner. J. Leukoc. Biol. 91, 967–976 (2012).

   Google Scholar 

7. Wahab, S. et al. (Licorice): A comprehensive review on its phytochemistry, biological activities, clinical evidence and toxicology. Plants 10 https://doi.org/10.3390/plants10122751 (2021).

8. Yang, R., Yuan, B. C., Ma, Y. S., Zhou, S. & Liu, Y. The anti-inflammatory activity of licorice, a widely used Chinese herb. Pharm. Biol. 55, 5–18 (2017).

   Google Scholar 

9. Sharifi-Rad, J. et al. Genus: enlightening phytochemical components for pharmacological and health-promoting abilities. Oxid. Med Cell Longev. 2021, 7571132 (2021).

   Google Scholar 

10. Mollica, L. et al. Glycyrrhizin binds to high-mobility group box 1 protein and inhibits its cytokine activities. Chem. Biol. 14, 431–441 (2007).

    Google Scholar 

11. Zhai, K. F. et al. Liquiritin from Glycyrrhiza uralensis Attenuating Rheumatoid Arthritis via Reducing Inflammation, Suppressing Angiogenesis, and Inhibiting MAPK Signaling Pathway. J. Agric Food Chem. 67, 2856–2864 (2019).

    Google Scholar 

12. Li, M. et al. Anti-inflammation of isoliquiritigenin via the inhibition of NF-κB and MAPK in LPS-stimulated MAC-T cells. BMC Vet. Res. 18, 320 (2022).

    Google Scholar 

13. Zhao, F. et al. Glycyrrhizin protects rats from sepsis by blocking HMGB1 signaling. Biomed. Res. Int. 2017, 9719647 (2017).

    Google Scholar 

14. Kim, Y. W. et al. Anti-inflammatory effects of liquiritigenin as a consequence of the inhibition of NF-kappaB-dependent iNOS and proinflammatory cytokines production. Br. J. Pharm. 154, 165–173 (2008).

    Google Scholar 

15. Zhang, Q. H. et al. Traditional uses, pharmacological effects, and molecular mechanisms of licorice in potential therapy of COVID-19. Front. Pharm. 12, 719758 (2021).

    Google Scholar 

16. Sharma, K. et al. Ultradeep human phosphoproteome reveals a distinct regulatory nature of Tyr and Ser/Thr-based signaling. Cell Rep. 8, 1583–1594 (2014).

    Google Scholar 

17. Kiuchi, S. et al. Using variable data-independent acquisition for capillary electrophoresis-based untargeted metabolomics. J. Am. Soc. Mass Spectrom. 35, 2118–2127 (2024).

    Google Scholar 

18. Kaneko, A. et al. Glucuronides of phytoestrogen flavonoid enhance macrophage function via conversion to aglycones by beta-glucuronidase in macrophages. Immun. Inflamm. Dis. 5, 265–279 (2017).

    Google Scholar 

19. Rodríguez-Prados, J. C. et al. Substrate fate in activated macrophages: a comparison between innate, classic, and alternative activation. J. Immunol. 185, 605–614 (2010).

    Google Scholar 

20. Sanchez-Lopez, E. et al. Choline uptake and metabolism modulate macrophage IL-1β and IL-18 production. Cell Metab. 29, 1350–1362.e1357 (2019).

    Google Scholar 

21. Lampropoulou, V. et al. Itaconate links inhibition of succinate dehydrogenase with macrophage metabolic remodeling and regulation of inflammation. Cell Metab. 24, 158–166 (2016).

    Google Scholar 

22. Marrocco, A. & Ortiz, L. A. Role of metabolic reprogramming in pro-inflammatory cytokine secretion from LPS or silica-activated macrophages. Front. Immunol. 13, 936167 (2022).

    Google Scholar 

23. Palmieri, E. M. et al. Nitric oxide orchestrates metabolic rewiring in M1 macrophages by targeting aconitase 2 and pyruvate dehydrogenase. Nat. Commun. 11, 698 (2020).

    Google Scholar 

24. Funk, J. L., Feingold, K. R., Moser, A. H. & Grunfeld, C. Lipopolysaccharide stimulation of RAW 264.7 macrophages induces lipid accumulation and foam cell formation. Atherosclerosis 98, 67–82 (1993).

    Google Scholar 

25. Huang, Y. L. et al. Toll-like receptor agonists promote prolonged triglyceride storage in macrophages. J. Biol. Chem. 289, 3001–3012 (2014).

    Google Scholar 

26. Castoldi, A. et al. Triacylglycerol synthesis enhances macrophage inflammatory function. Nat. Commun. 11, 4107 (2020).

    Google Scholar 

27. Kim, S. J. et al. AMPK phosphorylates Desnutrin/ATGL and hormone-sensitive lipase to regulate lipolysis and fatty acid oxidation within adipose tissue. Mol. Cell Biol. 36, 1961–1976 (2016).

    Google Scholar 

28. Bekker-Jensen, D. B. et al. Rapid and site-specific deep phosphoproteome profiling by data-independent acquisition without the need for spectral libraries. Nat. Commun. 11, 787 (2020).

    Google Scholar 

29. Katsuyuki, Y. & Shinya, K. Metabolism as a signal generator across trans-omic networks at distinct time scales. Curr. Opin. Syst. Biol. 8, 59–66 (2018).

    Google Scholar 

30. Kanehisa, M., Furumichi, M., Sato, Y., Matsuura, Y. & Ishiguro-Watanabe, M. KEGG: biological systems database as a model of the real world. Nucleic Acids Res. 53, D672–D677 (2025).

    Google Scholar 

31. Chen, C. et al. Suppression of lung cancer progression by isoliquiritigenin through its metabolite 2, 4, 2’, 4’-Tetrahydroxychalcone. J. Exp. Clin. Cancer Res. 37, 243 (2018).

    Google Scholar 

32. Van Nostrand, J. L. et al. AMPK regulation of Raptor and TSC2 mediate metformin effects on transcriptional control of anabolism and inflammation. Genes Dev. 34, 1330–1344 (2020).

    Google Scholar 

33. Yamamoto, H. Multiset partial least squares with rank order of groups for integrating multi-omics data. Preprint at https://doi.org/10.1101/2022.08.30.505949 (2022).

34. Yamamoto, H. PLS-ROG: partial least squares with rank order of groups. J. Chemometr. 31 https://doi.org/10.1002/cem.2883 (2017).

35. Argelaguet, R. et al. Multi-omics factor analysis-a framework for unsupervised integration of multi-omics data sets. Mol. Syst. Biol. 14 https://doi.org/10.15252/msb.20178124 (2018).

36. Argelaguet, R. et al. MOFA plus: a statistical framework for comprehensive integration of multi-modal single-cell data. Genome Biol. 21 https://doi.org/10.1186/s13059-020-02015-1 (2020).

37. Lu, W., Ji, H. & Wu, D. SIRT2 plays complex roles in neuroinflammation neuroimmunology-associated disorders. Front. Immunol. 14, 1174180 (2023).

    Google Scholar 

38. Pandithage, R. et al. The regulation of SIRT2 function by cyclin-dependent kinases affects cell motility. J. Cell Biol. 180, 915–929 (2008).

    Google Scholar 

39. Zhao, X. et al. Sirt1 inhibits macrophage polarization and inflammation in gouty arthritis by inhibiting the MAPK/NF-κB/AP-1 pathway and activating the Nrf2/HO-1 pathway. Inflamm. Res. 73, 1173–1184 (2024).

    Google Scholar 

40. Xia, Y. et al. GABA transporter sustains IL-1β production in macrophages. Sci. Adv. 7 https://doi.org/10.1126/sciadv.abe9274 (2021).

41. Kim, J. Y. et al. Isoliquiritigenin isolated from the roots of Glycyrrhiza uralensis inhibits LPS-induced iNOS and COX-2 expression via the attenuation of NF-kappaB in RAW 264.7 macrophages. Eur. J. Pharm. 584, 175–184 (2008).

    Google Scholar 

42. Mills, E. L. et al. Itaconate is an anti-inflammatory metabolite that activates Nrf2 via alkylation of KEAP1. Nature 556, 113–117 (2018).

    Google Scholar 

43. Yang, H. Y. et al. SIRT1 Activators Suppress Inflammatory Responses through Promotion of p65 Deacetylation and Inhibition of NF-κB Activity. Plos ONE 7 https://doi.org/10.1371/journal.pone.0046364 (2012).

44. Rothgiesser, K. M., Erener, S., Waibel, S., Lüscher, B. & Hottiger, M. O. SIRT2 regulates NF-κB-dependent gene expression through deacetylation of p65 Lys310. J. Cell Sci. 123, 4251–4258 (2010).

    Google Scholar 

45. Tao, Z. J., Jin, Z. H., Wu, J. B., Cai, G. J. & Yu, X. L. Sirtuin family in autoimmune diseases. Front. Immunol. 14 https://doi.org/10.3389/fimmu.2023.1186231 (2023).

46. Ruderman, N. B. et al. AMPK and SIRT1: a long-standing partnership?. Am. J. Physiol. Endocrinol. Metab. 298, E751–E760 (2010).

    Google Scholar 

47. Wang, H. W. et al. Blockade of fatty acid signalling inhibits lipopolysaccharide-induced macrophage recruitment and progression of apical periodontitis. Int. Endod. J. 54, 902–915 (2021).

    Google Scholar 

48. Xiao, Y., Yang, Y., Xiong, H. & Dong, G. The implications of FASN in immune cell biology and related diseases. Cell Death Dis. 15, 88 (2024).

    Google Scholar 

49. Tzou, F. Y., Hornemann, T., Yeh, J. Y. & Huang, S. Y. The pathophysiological role of dihydroceramide desaturase in the nervous system. Prog. Lipid Res. 91, 101236 (2023).

    Google Scholar 

50. Bhandage, A. K. & Barragan, A. GABAergic signaling by cells of the immune system: more the rule than the exception. Cell Mol. Life Sci. 78, 5667–5679 (2021).

    Google Scholar 

51. Jin, Z., Mendu, S. K. & Birnir, B. GABA is an effective immunomodulatory molecule. Amino Acids 45, 87–94 (2013).

    Google Scholar 

52. Tang, D., Orlandi, P., Li, Q., Bandini, A. & Bocci, G. GABAergic signaling in colorectal cancer: mechanistic insights, tumor microenvironment crosstalk, and therapeutic opportunities. Biochim. Biophys. Acta Rev. Cancer 1880, 189414 (2025).

    Google Scholar 

53. Ortega, A. A new role for GABA: inhibition of tumor cell migration. Trends Pharm. Sci. 24, 151–154 (2003).

    Google Scholar 

54. Eid, H. M. A. et al. GABA and GABAergic dysfunction in COVID-19: Piecing the puzzle with targeting immunity and several inflammatory pathways. Cytokine 193 https://doi.org/10.1016/j.cyto.2025.156976 (2025).

55. Zhang, Q. L. et al. Insights and progress on the biosynthesis, metabolism, and physiological functions of gamma-aminobutyric acid (GABA): a review. Peerj 12 https://doi.org/10.7717/peerj.18712 (2024).

56. Schildberger, A., Rossmanith, E., Eichhorn, T., Strassl, K. & Weber, V. Monocytes, peripheral blood mononuclear cells, and THP-1 cells exhibit different cytokine expression patterns following stimulation with lipopolysaccharide. Mediat. Inflamm. 2013 https://doi.org/10.1155/2013/697972 (2013).

57. Trost, M. et al. The Phagosomal Proteome in Interferon-γ-Activated Macrophages. Immunity 30, 143–154 (2009).

    Google Scholar 

58. Sun, Y., Mehmood, A., Battino, M., Xiao, J. B. & Chen, X. M. Enrichment of gamma-aminobutyric acid in foods: From conventional methods to innovative technologies. Food Res. Int. 162 https://doi.org/10.1016/j.foodres.2022.111801 (2022).

59. Sasaki, K. et al. Metabolomics platform with capillary electrophoresis coupled with high-resolution mass spectrometry for plasma analysis. Anal. Chem. 91, 1295–1301 (2019).

    Google Scholar 

60. Takeda, H. et al. MS-DIAL 5 multimodal mass spectrometry data mining unveils lipidome complexities. Nat. Commun. 15, 9903 (2024).

    Google Scholar 

61. BLIGH, E. G. & DYER, W. J. A rapid method of total lipid extraction and purification. Can. J. Biochem. Physiol. 37, 911–917 (1959).

    Google Scholar 

62. Tsugawa, H. et al. A lipidome landscape of aging in mice. Nat. Aging 4, 709–726 (2024).

    Google Scholar 

63. Masuda, T., Tomita, M. & Ishihama, Y. Phase transfer surfactant-aided trypsin digestion for membrane proteome analysis. J. Proteome Res. 7, 731–740 (2008).

    Google Scholar 

64. Nakagami, H. StageTip-based HAMMOC, an efficient and inexpensive phosphopeptide enrichment method for plant shotgun phosphoproteomics. Methods Mol. Biol. 1072, 595–607 (2014).

    Google Scholar 

65. Sugiyama, N. et al. Phosphopeptide enrichment by aliphatic hydroxy acid-modified metal oxide chromatography for nano-LC-MS/MS in proteomics applications. Mol. Cell Proteom. 6, 1103–1109 (2007).

    Google Scholar 

66. Rappsilber, J., Mann, M. & Ishihama, Y. Protocol for micro-purification, enrichment, pre-fractionation and storage of peptides for proteomics using StageTips. Nat. Protoc. 2, 1896–1906 (2007).

    Google Scholar 

67. Kong, A. T., Leprevost, F. V., Avtonomov, D. M., Mellacheruvu, D. & Nesvizhskii, A. I. MSFragger: ultrafast and comprehensive peptide identification in mass spectrometry-based proteomics. Nat. Methods 14, 513–520 (2017).

    Google Scholar 

68. da Veiga Leprevost, F. et al. Philosopher: a versatile toolkit for shotgun proteomics data analysis. Nat. Methods 17, 869–870 (2020).

    Google Scholar 

69. Yu, F., Haynes, S. E. & Nesvizhskii, A. I. IonQuant enables accurate and sensitive label-free quantification with FDR-controlled match-between-runs. Mol. Cell Proteom. 20, 100077 (2021).

    Google Scholar 

70. Pang, Z. Q. et al. MetaboAnalyst 6.0: towards a unified platform for metabolomics data processing, analysis and interpretation. Nucleic Acids Res. 52, W398–W406 (2024).

    Google Scholar 

Download references