Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Article
  • Published:

Techniques and Methods

Butyrate and tributyrin reduce LPS-induced inflammatory cytokine production from human visceral fat

International Journal of Obesity volume 48, pages 1559–1567 (2024)Cite this article

Subjects

Abstract

Introduction

The current obesity crisis has resulted in many people with excess adipose tissue suffering from chronic inflammation. This inflammation is largely due to the release of cytokines and chemokines from visceral fat. The aim of this study was to identify potential anti-inflammatory agents that might alleviate obesity-induced chronic inflammation.

Methods

To identify agents that might alleviate this obesity-induced chronic inflammation we have developed a simple protocol for incubating intact pieces of human visceral adipose tissue in 35 mm tissue culture plates, in the presence of low-dose lipopolysaccharide (LPS) and co-incubating these samples with potential anti-inflammatory agents. RNA-Seq analysis was performed to identify enriched gene expression signatures among the most significantly differentially expressed genes.

Results

From this screen, we have identified the short-chain fatty acid (SCFA) sodium butyrate and its triacylglyceride form, tributyrin, as effective agents, significantly reducing the production of LPS-induced inflammatory cytokines and chemokines from all adipose tissue samples tested. As well, these agents appear to be non-toxic at the concentrations tested. RNA-Seq analysis has revealed that IL36γ is one of the most upregulated genes in response to LPS and one of the most downregulated when sodium butyrate is added to human fat samples stimulated with LPS. IL-36γ ELISAs confirmed this holds true at the protein level as well.

Conclusions

These studies suggest that the short-chain fatty acid, sodium butyrate, and its triacylglyceride form, tributyrin, might alleviate the chronic inflammation that is associated with many individuals with obesity.

Access through your institution
Buy or subscribe

This is a preview of subscription content, access via your institution

Access options

Access through your institution

Buy this article

  • Purchase on SpringerLink
  • Instant access to full article PDF
Buy now

Prices may be subject to local taxes which are calculated during checkout

Fig. 1: Development of the Ex-Vivo Visceral Fat Assay.
Fig. 2: Optimization of the Ex-Vivo Visceral Fat Assay.
Fig. 3: Ex-vivo Visceral Fat Screen Identifies Sodium Butyrate and Tributyrin as Potential Anti-Inflammatory Agents.
Fig. 4: Sodium Butyrate and Tributyrin Suppress LPS-induced Cytokines/Chemokines from Visceral Fat.
Fig. 5: RNA-seq analysis of intact visceral fat pieces stimulated or not with LPS ± sodium butyrate.

Similar content being viewed by others

Data availability

All data relevant to the study are included in the article.

References

  1. Huttunen R, Syrjanen J. Obesity and the risk and outcome of infection. Int J Obes. 2013;37:333–40.

    Article  CAS  Google Scholar 

  2. Honce R, Schultz-Cherry S. Impact of obesity on influenza A virus pathogenesis, immune response, and evolution. Front Immunol. 2019;10:1071.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  3. Deng T, Lyon CJ, Bergin S, Caligiuri MA, Hsueh WA. Obesity, inflammation, and cancer. Annu Rev Pathol. 2016;11:421–49.

    Article  CAS  PubMed  Google Scholar 

  4. Lee H. Obesity-associated cancers: evidence from studies in mouse models. Cells. 2022;11:1472.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  5. Barton JR, Snook AE, Waldman SA. From leptin to lasers: the past and present of mouse models of obesity. Expert Opin Drug Discov. 2021;16:777–90.

    Article  PubMed  PubMed Central  Google Scholar 

  6. Rana MN, Neeland IJ. Adipose tissue inflammation and cardiovascular disease: an update. Curr Diab Rep. 2022;22:27–37.

    Article  CAS  PubMed  Google Scholar 

  7. Atkuri KR, Herzenberg LA, Niemi AK, Cowan T, Herzenberg LA. Importance of culturing primary lymphocytes at physiological oxygen levels. Proc Natl Acad Sci USA. 2007;104:4547–52.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  8. Wu D, Han JM, Yu X, Lam AJ, Hoeppli RE, Pesenacker AM, et al. Characterization of regulatory T cells in obese omental adipose tissue in humans. Eur J Immunol. 2019;49:336–47.

    Article  CAS  PubMed  Google Scholar 

  9. Carswell KA, Lee MJ, Fried SK. Culture of isolated human adipocytes and isolated adipose tissue. Methods Mol Biol. 2012;806:203–14.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  10. Dobin A, Davis CA, Schlesinger F, Drenkow J, Zaleski C, Jha S, et al. STAR: ultrafast universal RNA-seq aligner. Bioinformatics. 2013;29:15–21.

    Article  CAS  PubMed  Google Scholar 

  11. Robinson MD, McCarthy DJ, Smyth GK. edgeR: a Bioconductor package for differential expression analysis of digital gene expression data. Bioinformatics. 2010;26:139–40.

    Article  CAS  PubMed  Google Scholar 

  12. Ritchie ME, Phipson B, Wu D, Hu Y, Law CW, Shi W, et al. limma powers differential expression analyses for RNA-sequencing and microarray studies. Nucleic Acids Res. 2015;43:e47.

    Article  PubMed  PubMed Central  Google Scholar 

  13. Yu G, Wang LG, Han Y, He QY. clusterProfiler: an R package for comparing biological themes among gene clusters. Omics. 2012;16:284–7.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  14. Kolb H. Obese visceral fat tissue inflammation: from protective to detrimental? BMC Med. 2022;20:494.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  15. Višnjić D, Lalić H, Dembitz V, Tomić B, Smoljo T. AICAr, a widely used AMPK activator with important AMPK-independent effects: a systematic review. Cells. 2021;10:1095.

    Article  PubMed  PubMed Central  Google Scholar 

  16. Elisia I, Lam V, Cho B, Hay M, Li MY, Kapeluto J, et al. Exploratory examination of inflammation state, immune response and blood cell composition in a human obese cohort to identify potential markers predicting cancer risk. PLoS ONE. 2020;15:e0228633.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  17. Zatterale F, Longo M, Naderi J, Raciti GA, Desiderio A, Miele C, et al. Chronic adipose tissue inflammation linking obesity to insulin resistance and type 2 diabetes. Front Physiol. 2020;10:1607.

    Article  PubMed  PubMed Central  Google Scholar 

  18. Kawai T, Autieri MV, Scalia R. Adipose tissue inflammation and metabolic dysfunction in obesity. Am J Physiol Cell Physiol. 2021;320:C375–91.

    Article  CAS  PubMed  Google Scholar 

  19. Weisberg SP, McCann D, Desai M, Rosenbaum M, Leibel RL, Ferrante AW Jr. Obesity is associated with macrophage accumulation in adipose tissue. J Clin Invest. 2003;112:1796–808.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  20. Shuster A, Patlas M, Pinthus JH, Mourtzakis M. The clinical importance of visceral adiposity: a critical review of methods for visceral adipose tissue analysis. Br J Radio. 2012;85:1–10.

    Article  CAS  Google Scholar 

  21. Saltiel AR, Olefsky JM. Inflammatory mechanisms linking obesity and metabolic disease. J Clin Invest. 2017;127:1–4.

    Article  PubMed  PubMed Central  Google Scholar 

  22. van Galen KA, Schrantee A, ter Horst KW, la Fleur SE, Booij J, Constable RT, et al. Brain responses to nutrients are severely impaired and not reversed by weight loss in humans with obesity: a randomized crossover study. Nat Metab. 2023;5:1059–72.

    Article  PubMed  Google Scholar 

  23. Hall KD, Kahan S. Maintenance of lost weight and long-term management of obesity. Med Clin North Am. 2018;102:183–97.

    Article  PubMed  PubMed Central  Google Scholar 

  24. van Deuren T, Blaak EE, Canfora EE. Butyrate to combat obesity and obesity-associated metabolic disorders: current status and future implications for therapeutic use. Obes Rev. 2022;23:e13498.

    Article  PubMed  PubMed Central  Google Scholar 

  25. Elisia I, Krystal G. The pros and cons of low carbohydrate and ketogenic diets in the prevention and treatment of cancer. Front Nutr. 2021;8:634845.

    Article  PubMed  PubMed Central  Google Scholar 

  26. Frühbeck G, Gómez-Ambrosi J, Ramírez B, Mentxaka A, Rodríguez A, Becerril S, et al. Increased levels of interleukin-36 in obesity and type 2 diabetes fuel adipose tissue inflammation by inducing its own expression and release by adipocytes and macrophages. Front Immunol. 2022;13:832185.

    Article  PubMed  PubMed Central  Google Scholar 

  27. Elisia I, Pae HB, Lam V, Cederberg R, Hofs E, Krystal G. Comparison of RAW264.7, human whole blood and PBMC assays to screen for immunomodulators. J Immunol Methods. 2018;452:26–31.

    Article  CAS  PubMed  Google Scholar 

  28. Guo L-T, Wang S-Q, Su J, Xu L-X, Ji Z-Y, Zhang R-Y, et al. Baicalin ameliorates neuroinflammation-induced depressive-like behavior through inhibition of toll-like receptor 4 expression via the PI3K/AKT/FoxO1 pathway. J Neuroinflammation. 2019;16:95.

    Article  PubMed  PubMed Central  Google Scholar 

  29. Amini R, Yazdanparast R, Ghaffari SH. Apigenin modulates the expression levels of pro-inflammatory mediators to reduce the human insulin amyloid-induced oxidant damages in SK-N-MC cells. Hum Exp Toxicol. 2014;34:642–53.

    Article  PubMed  Google Scholar 

  30. Sakai C, Ishida M, Ohba H, Yamashita H, Uchida H, Yoshizumi M, et al. Fish oil omega-3 polyunsaturated fatty acids attenuate oxidative stress-induced DNA damage in vascular endothelial cells. PLoS ONE. 2017;12:e0187934.

    Article  PubMed  PubMed Central  Google Scholar 

  31. Ayasolla KR, Singh AK, Singh I. 5-aminoimidazole-4-carboxamide-1-beta-4-ribofuranoside (AICAR) attenuates the expression of LPS- and Abeta peptide-induced inflammatory mediators in astroglia. J Neuroinflammation. 2005;2:21.

    Article  PubMed  PubMed Central  Google Scholar 

  32. Kim M, Nowakowski M, Joks R. Effects of curcumin on LPS-induced nitric oxide production from murine macrophages and PBMC of humans with respiratory allergic disease. J Allergy Clin Immunol. 2011;127:AB261.

    Article  Google Scholar 

  33. LaMoia TE, Shulman GI. Cellular and molecular mechanisms of metformin action. Endocr Rev. 2020;42:77–96.

    Article  PubMed Central  Google Scholar 

  34. Hoppe J, Schäfer R, Hoppe V, Sachinidis A. ATP and adenosine prevent via different pathways the activation of caspases in apoptotic AKR-2B fibroblasts. Cell Death Differ. 1999;6:546–56.

    Article  CAS  PubMed  Google Scholar 

  35. Rafiei H, Yeung M, Kowalski S, Krystal G, Elisia I. Development of a novel human triculture model of non-alcoholic fatty liver disease and identification of berberine as ameliorating steatosis, oxidative stress and fibrosis. Front Pharmacol. 2023;14:1234300.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  36. Elisia I, Kitts DD. Different tocopherol isoforms vary in capacity to scavenge free radicals, prevent inflammatory response, and induce apoptosis in both adult- and fetal-derived intestinal epithelial cells. Biofactors. 2013;39:663–71.

    Article  CAS  PubMed  Google Scholar 

  37. Elisia I, Kitts DD. Tocopherol isoforms (alpha-, gamma-, and delta-) show distinct capacities to control Nrf-2 and NfκB signaling pathways that modulate inflammatory response in Caco-2 intestinal cells. Mol Cell Biochem. 2015;404:123–31.

    Article  CAS  PubMed  Google Scholar 

  38. Elisia I, Kitts DD. Modulation of NF-κB and Nrf2 control of inflammatory responses in FHs 74 Int cell line is tocopherol isoform-specific. Am J Physiol Gastrointest Liver Physiol. 2013;305:G940–9.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  39. Kieronska-Rudek A, Kij A, Kaczara P, Tworzydlo A, Napiorkowski M, Sidoryk K. et al. Exogenous vitamins K exert anti-inflammatory effects dissociated from their role as substrates for synthesis of endogenous MK-4 in murine macrophages cell line. Cells. 2021;10:1571.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  40. Tuohimaa P, Golovko O, Kalueff A, Nazarova N, Qiao S, Syvälä H, et al. Calcidiol and prostate cancer. J Steroid Biochem Mol Biol. 2005;93:183–90.

    Article  CAS  PubMed  Google Scholar 

  41. Wang J, Guo S, Xu X, Zhang C. 1,25-Dihydroxyvitamin D3: a positive factor for the osteogenic differentiation of hPDLSCs and for the tissue regenerative activity of cell sheets. Cell Transpl. 2023;32:09636897231202541.

    Article  Google Scholar 

  42. Thai PN, Miller CV, King MT, Schaefer S, Veech RL, Chiamvimonvat N, et al. Ketone ester D‐β‐hydroxybutyrate‐(R)‐1,3 butanediol prevents decline in cardiac function in type 2 diabetic mice. J Am Heart Assoc. 2021;10:e020729.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  43. Mazurek S, Michel A, Eigenbrodt E. Effect of extracellular AMP on cell proliferation and metabolism of breast cancer cell lines with high and low glycolytic rates. J Biol Chem. 1997;272:4941–52.

    Article  CAS  PubMed  Google Scholar 

  44. Adzic M, Stevanovic I, Josipovic N, Laketa D, Lavrnja I, Bjelobaba IM, et al. Extracellular ATP induces graded reactive response of astrocytes and strengthens their antioxidative defense in vitro. J Neurosci Res. 2017;95:1053–66.

    Article  CAS  PubMed  Google Scholar 

  45. Hidalgo M, Martin-Santamaria S, Recio I, Sanchez-Moreno C, de Pascual-Teresa B, Rimbach G, et al. Potential anti-inflammatory, anti-adhesive, anti/estrogenic, and angiotensin-converting enzyme inhibitory activities of anthocyanins and their gut metabolites. Genes Nutr. 2012;7:295–306.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  46. Winter AN, Brenner MC, Punessen N, Snodgrass M, Byars C, Arora Y, et al. Comparison of the neuroprotective and anti-inflammatory effects of the anthocyanin metabolites, protocatechuic acid and 4-hydroxybenzoic acid. Oxid Med Cell Longev. 2017;2017:6297080.

    Article  PubMed  PubMed Central  Google Scholar 

  47. Walker J, Schueller K, Schaefer L-M, Pignitter M, Esefelder L, Somoza V. Resveratrol and its metabolites inhibit pro-inflammatory effects of lipopolysaccharides in U-937 macrophages in plasma-representative concentrations. Food Func. 2014;5:74–84.

    Article  CAS  Google Scholar 

  48. Kondreddy VKR, Kamatham AN. Celecoxib, a COX-2 inhibitor, synergistically potentiates the anti-inflammatory activity of docosahexaenoic acid in macrophage cell line. Immunopharmacol Immunotoxicol. 2016;38:153–61.

    Article  CAS  PubMed  Google Scholar 

  49. Kawase T, Orikasa M, Suzuki A. Effects of prostaglandin E2 and F2α on cytoplasmic pH in a clonal osteoblast-like cell line, MOB 3–4. J Cell Phys. 1991;146:141–7.

    Article  CAS  Google Scholar 

  50. Chen Q, Ross AC. Retinoic acid regulates cell cycle progression and cell differentiation in human monocytic THP-1 cells. Exp Cell Res. 2004;297:68–81.

    Article  CAS  PubMed  Google Scholar 

Download references

Acknowledgements

The research was supported by the Lotte and John Hecht Memorial Foundation (#3693), with core support from the BC Cancer Foundation and BC Cancer. This study was in part supported by Program Project Grant funding from the Terry Fox Research Institute (Grant No. 1108 to CS). MYL is a PhD candidate at the University of British Columbia, Canada, and is supported by an Elizabeth C. Watters Research Fellowship.

Author information

Author notes

  1. These authors contributed equally: Hossein Rafiei, Michelle Yeung, Sara Kowalski.

Authors and Affiliations

  1. Terry Fox Laboratory, BC Cancer Research Institute, Vancouver, BC, V5Z 1L3, Canada

    Hossein Rafiei, Michelle Yeung, Sara Kowalski, Serena Hollman, Gerald Krystal & Ingrid Elisia

  2. Department of Lymphoid Cancer Research, BC Cancer Research Institute, Vancouver, BC, V5Z 1L3, Canada

    Michael Yu Li, Gerben Duns, Luke O’Brien & Christian Steidl

  3. Department of Pathology and Laboratory Medicine, University of British Columbia, Vancouver, BC, V5Z 1L3, Canada

    Michael Yu Li & Christian Steidl

  4. Richmond Metabolic and Bariatric Surgery Program, Richmond Hospital, Vancouver Coastal Health, Richmond, BC, V7C 5L9, Canada

    David Harris, Jacqueline Chang, Nam Nguyen, Ekua Yorke & Sharadh Sampath

Authors
  1. Hossein Rafiei
    View author publications

    Search author on:PubMed Google Scholar

  2. Michelle Yeung
    View author publications

    Search author on:PubMed Google Scholar

  3. Sara Kowalski
    View author publications

    Search author on:PubMed Google Scholar

  4. Michael Yu Li
    View author publications

    Search author on:PubMed Google Scholar

  5. David Harris
    View author publications

    Search author on:PubMed Google Scholar

  6. Jacqueline Chang
    View author publications

    Search author on:PubMed Google Scholar

  7. Nam Nguyen
    View author publications

    Search author on:PubMed Google Scholar

  8. Ekua Yorke
    View author publications

    Search author on:PubMed Google Scholar

  9. Sharadh Sampath
    View author publications

    Search author on:PubMed Google Scholar

  10. Serena Hollman
    View author publications

    Search author on:PubMed Google Scholar

  11. Gerben Duns
    View author publications

    Search author on:PubMed Google Scholar

  12. Luke O’Brien
    View author publications

    Search author on:PubMed Google Scholar

  13. Christian Steidl
    View author publications

    Search author on:PubMed Google Scholar

  14. Gerald Krystal
    View author publications

    Search author on:PubMed Google Scholar

  15. Ingrid Elisia
    View author publications

    Search author on:PubMed Google Scholar

Contributions

IE, GK, and HR conceptualized the study; IE, HR, MY, SK, MYL, SH, and LB performed experiments; IE, HR, MY, SK, SH, and GD analyzed data and figures; IE, MYL, GD, and GK wrote the first draft; IE, HR, MY, MYL, GD, DH, CS, and GK reviewed and revised the manuscript; NN, EY, and SS performed the bariatric surgeries; DH, JC, CS, and GK provided resources and supervision; all authors have read and agreed to the final version of the manuscript.

Corresponding author

Correspondence to Ingrid Elisia.

Ethics declarations

Competing interests

CS has performed consultancy for Bayer and Eisai, and has received research funding from Epizyme and Trillium Therapeutics.

Additional information

Publisher’s note Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Supplementary information

Rights and permissions

Springer Nature or its licensor (e.g. a society or other partner) holds exclusive rights to this article under a publishing agreement with the author(s) or other rightsholder(s); author self-archiving of the accepted manuscript version of this article is solely governed by the terms of such publishing agreement and applicable law.

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Rafiei, H., Yeung, M., Kowalski, S. et al. Butyrate and tributyrin reduce LPS-induced inflammatory cytokine production from human visceral fat. Int J Obes 48, 1559–1567 (2024). https://doi.org/10.1038/s41366-024-01581-9

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue date:

  • DOI: https://doi.org/10.1038/s41366-024-01581-9

Search

Advanced search

Quick links