Dietary Omega-3 Supplementation Shapes Gut Microbiota and Regulates Immunometabolism in a Mouse Model of Obesity
Main Article Content
Abstract
Host-gut microbiota interactions play a pivotal role in shaping the delicate balance between health and disease within the human body. The impact of dietary factors, specifically high fat content diets on gut microbiota composition has been widely demonstrated. We have previously shown that the constant and sustained administration of Omega-3 fatty acids induced specific changes in gut microbiota composition, modulating the immune metabolic response of visceral adipose tissue in our mouse model of obesity. We now set out to determine if this effect is Omega-3 dose-dependent. To achieve this, C57BL/6J(B6) mice were fed for 24 weeks with three diets, two with medium content total fat, but different Omega-3 content and a control diet.
Gut microbiota composition, metabolic biomarkers and immune cells in visceral adipose tissue were analyzed. A distinctive segregation of gut microbiota composition, a significantly higher proportion of regulatory T cells (CD45+CD4+ FoxP3+), Omega-3 dose dependent and increased levels of leptin and cholesterol with no differences in adiponectin values were found in fat fed groups. Simple mediation analyses revealed significant associations between the microbial profile and immunometabolic regulation. To remark, is the capacity of Lachnospiraceae UCG-001 to modulate levels of leptin, glucose, and cholesterol through the stimulation of CD45+CD4+FOXP3+IL10+ cells. Our findings suggest a modulatory effect of omega-3 fatty acids on the microbiota, the metabolism, and the immunoregulatory capacity of visceral adipose tissue, supporting the hypothesis that alteration of the gut microbiota composition by omega-3 fatty acids may be a promising approach in managing obesity and associated metabolic diseases.
Keywords:
obesity, immune-metabolism, adipose tissue regulatory T cells, gut microbiota; Omega-3, high fat diet, visceral adipose tissue, regulatory T cells
Article Details
How to Cite
PORTELA, Néstor D. et al.
Dietary Omega-3 Supplementation Shapes Gut Microbiota and Regulates Immunometabolism in a Mouse Model of Obesity.
Medical Research Archives, [S.l.], v. 13, n. 5, may 2025.
ISSN 2375-1924.
Available at: <https://esmed.org/MRA/mra/article/view/6623>. Date accessed: 20 june 2025.
doi: https://doi.org/10.18103/mra.v13i5.6623.
Section
Research Articles
The Medical Research Archives grants authors the right to publish and reproduce the unrevised contribution in whole or in part at any time and in any form for any scholarly non-commercial purpose with the condition that all publications of the contribution include a full citation to the journal as published by the Medical Research Archives.
References
1. Bray GA, Kim KK, Wilding JPH, World Obesity Federation. Obesity: a chronic relapsing progressive disease process. A position statement of the World Obesity Federation. Obes Rev. 2017;18(7):715-723. doi:10.1111/obr.12551
2. Schwartz MW, Seeley RJ, Zeltser LM, et al. Obesity Pathogenesis: An Endocrine Society Scientific Statement. Endocr Rev. 2017;38(4):267-296. doi:10.1210/er.2017-00111
3. Xu H, Barnes GT, Yang Q, et al. Chronic inflammation in fat plays a crucial role in the development of obesity-related insulin resistance. J Clin Invest. 2003;112(12):1821-1830. doi:10.1172/JCI200319451
4. Ley RE, Bäckhed F, Turnbaugh P, Lozupone CA, Knight RD, Gordon JI. Obesity alters gut microbial ecology. Proc Natl Acad Sci U S A. 2005;102(31):11070-11075. doi:10.1073/pnas.0504978102
5. Ley RE, Turnbaugh PJ, Klein S, Gordon JI. Human gut microbes associated with obesity. Nature. 2006;444(7122):1022-1023. doi:10.1038/4441022a
6. Le Chatelier E, Nielsen T, Qin J, et al. Richness of human gut microbiome correlates with metabolic markers. Nature. 2013;500(7464):541-546. doi:10.1038/nature12506
7. Busebee B, Ghusn W, Cifuentes L, Acosta A. Obesity: A Review of Pathophysiology and Classification. Mayo Clinic Proceedings. 2023;98(12):1842-1857. doi:10.1016/j.mayocp.2023.05.026
8. González-Muniesa P, Mártinez-González MA, Hu FB, et al. Obesity. Nat Rev Dis Primers. 2017;3(1):17034. doi:10.1038/nrdp.2017.34
9. Castoldi A, Naffah de Souza C, Câmara NOS, Moraes-Vieira PM. The Macrophage Switch in Obesity Development. Front Immunol. 2016;6:637. doi:10.3389/fimmu.2015.00637
10. Zatterale F, Longo M, Naderi J, et al. Chronic Adipose Tissue Inflammation Linking Obesity to Insulin Resistance and Type 2 Diabetes. Front Physiol. 2020;10:1607. doi:10.3389/fphys.2019.01607
11. Nishimura S, Manabe I, Nagasaki M, et al. CD8+ effector T cells contribute to macrophage recruitment and adipose tissue inflammation in obesity. Nat Med. 2009;15(8):914-920. doi:10.1038/nm.1964
12. Weisberg SP, McCann D, Desai M, Rosenbaum M, Leibel RL, Ferrante AW. Obesity is associated with macrophage accumulation in adipose tissue. J Clin Invest. 2003;112(12):1796-1808. doi:10.1172/JCI200319246
13. Bradley D, Deng T, Shantaram D, Hsueh WA. Orchestration of the Adipose Tissue Immune Landscape by Adipocytes. Published online February 12, 2024. doi:10.1146/annurev-physiol-042222-024353
14. Chait A, den Hartigh LJ. Adipose tissue distribution, inflammation and its metabolic consequences, including diabetes and cardiovascular disease. Front Cardiovasc Med. 2020;7:22. doi:10.3389/fcvm.2020.00022
15. Turnbaugh PJ, Ley RE, Mahowald MA, Magrini V, Mardis ER, Gordon JI. An obesity-associated gut microbiome with increased capacity for energy harvest. Nature. 2006;444(7122):1027-1031. doi:10.1038/nature05414
16. Cani PD, Amar J, Iglesias MA, et al. Metabolic endotoxemia initiates obesity and insulin resistance. Diabetes. 2007;56(7):1761-1772. doi:10.2337/db06-1491
17. Caesar R, Tremaroli V, Kovatcheva-Datchary P, Cani PD, Bäckhed F. Crosstalk between Gut Microbiota and Dietary Lipids Aggravates WAT Inflammation through TLR Signaling. Cell Metab. 2015;22(4):658-668. doi:10.1016/j.cmet.2015.07.026
18. Haeusler RA, Camastra S, Nannipieri M, et al. Increased Bile Acid Synthesis and Impaired Bile Acid Transport in Human Obesity. J Clin Endocrinol Metab. 2016;101(5):1935-1944. doi:10.1210/jc.2015-2583
19. Agans R, Gordon A, Kramer DL, Perez-Burillo S, Rufián-Henares JA, Paliy O. Dietary Fatty Acids Sustain the Growth of the Human Gut Microbiota. Appl Environ Microbiol. 2018;84(21):e01525-18. doi:10.1128/AEM.01525-18
20. Daniel H, Gholami AM, Berry D, et al. High-fat diet alters gut microbiota physiology in mice. ISME J. 2014;8(2):295-308. doi:10.1038/ismej.2013.155
21. Schoeler M, Caesar R. Dietary lipids, gut microbiota and lipid metabolism. Rev Endocr Metab Disord. 2019;20(4):461-472. doi:10.1007/s11154-019-09512-0
22. Xu AA, Kennedy LK, Hoffman K, et al. Dietary Fatty Acid Intake and the Colonic Gut Microbiota in Humans. Nutrients. 2022;14(13):2722. doi:10.3390/nu14132722
23. López-Millán E, Gutiérrez-Uribe JA, Antunes-Ricardo M. Omega-3 supplementation: Impact on low chronic inflammation associated with obesity. Trends in Food Science & Technology. 2025;155:104799. doi:10.1016/j.tifs.2024.104799
24. Menni C, Jackson MA, Pallister T, Steves CJ, Spector TD, Valdes AM. Gut microbiome diversity and high-fibre intake are related to lower long-term weight gain. Int J Obes. 2017;41(7):1099-1105. doi:10.1038/ijo.2017.66
25. Cao W, Liu F, Li RW, et al. Docosahexaenoic acid-rich fish oil prevented insulin resistance by modulating gut microbiome and promoting colonic peptide YY expression in diet-induced obesity mice. Food Science and Human Wellness. 2022;11(1):177-188. doi:10.1016/j.fshw.2021.07.018
26. Monk JM, Liddle DM, Hutchinson AL, et al. Fish oil supplementation to a high-fat diet improves both intestinal health and the systemic obese phenotype. The Journal of Nutritional Biochemistry. 2019;72:108216. doi:10.1016/j.jnutbio.2019.07.007
27. Watson H, Mitra S, Croden FC, et al. A randomised trial of the effect of omega-3 polyunsaturated fatty acid supplements on the human intestinal microbiota. Gut. 2018;67(11):1974-1983. doi:10.1136/gutjnl-2017-314968
28. Portela ND, Galván C, Sanmarco LM, et al. Omega-3-Supplemented Fat Diet Drives Immune Metabolic Response in Visceral Adipose Tissue by Modulating Gut Microbiota in a Mouse Model of Obesity. Nutrients. 2023;15(6):1404. doi:10.3390/nu15061404
29. Ellacott KLJ, Morton GJ, Woods SC, Tso P, Schwartz MW. Assessment of feeding behavior in laboratory mice. Cell Metab. 2010;12(1):10-17. doi:10.1016/j.cmet.2010.06.001
30. Callahan BJ, McMurdie PJ, Rosen MJ, Han AW, Johnson AJA, Holmes SP. DADA2: High resolution sample inference from Illumina amplicon data. Nat Methods. 2016;13(7):581-583. doi:10.1038/nmeth.3869
31. McMurdie PJ, Holmes S. phyloseq: An R Package for Reproducible Interactive Analysis and Graphics of Microbiome Census Data. PLOS ONE. 2013;8(4):e61217. doi:10.1371/journal.pone.0061217
32. Lahti L, Shetty S. microbiome R package. Published online 2017. doi:10.18129/B9.bioc.microbiome
33. R Core Team. R Core Team (2022). R: A language and environment for statistical computing. Published online 2021. https://www.R-project.org/.
34. Quast C, Pruesse E, Yilmaz P, et al. The SILVA ribosomal RNA gene database project: improved data processing and web-based tools. Nucleic Acids Research. 2013;41(D1):D590-D596. doi:10.1093/nar/gks1219
35. Cao Y, Dong Q, Wang D, Zhang P, Liu Y, Niu C. microbiomeMarker: an R/Bioconductor package for microbiome marker identification and visualization. Bioinformatics. 2022;38(16):4027-4029. doi:10.1093/bioinformatics/btac438
36. Curat CA, Miranville A, Sengenès C, et al. From Blood Monocytes to Adipose Tissue-Resident Macrophages: Induction of Diapedesis by Human Mature Adipocytes. Diabetes. 2004;53(5):1285-1292. doi:10.2337/diabetes.53.5.1285
37. Wei T SV. R package “corrplot”: Visualization of a Correlation Matrix. Published online 2021. https://github.com/taiyun/corrplot.
38. Wickham H. ggplot2: Elegant Graphics for Data Analysis. 2016. Accessed January 25, 2023. https://link.springer.com/book/10.1007/978-3-319-24277-4
39. Kassambara A. ggpubr: “ggplot2” Based Publication Ready Plots. R package version 0.5.0. Published online 2022. https://CRAN.R-project.org/package=ggpubr
40. Dixon P. VEGAN, A Package of R Functions for Community Ecology. Journal of Vegetation Science. 2003;14(6):927-930.
41. Rosseel Y, Jorgensen TD, Wilde LD, et al. lavaan: Latent Variable Analysis. Published online December 20, 2023. Accessed March 7, 2024. https://cran.r-project.org/web/packages/lavaan/index.html
42. Shen W, Wolf PG, Carbonero F, et al. Intestinal and systemic inflammatory responses are positively associated with sulfidogenic bacteria abundance in high-fat-fed male C57BL/6J mice. J Nutr. 2014;144(8):1181-1187. doi:10.3945/jn.114.194332
43. Hildebrandt MA, Hoffmann C, Sherrill-Mix SA, et al. High-fat diet determines the composition of the murine gut microbiome independently of obesity. Gastroenterology. 2009;137(5):1716-1724.e1-2. doi:10.1053/j.gastro.2009.08.042
44. LeMieux MJ, Kalupahana NS, Scoggin S, Moustaid-Moussa N. Eicosapentaenoic acid reduces adipocyte hypertrophy and inflammation in diet-induced obese mice in an adiposity-independent manner. J Nutr. 2015;145(3):411-417. doi:10.3945/jn.114.202952
45. Albracht-Schulte K, Kalupahana NS, Ramalingam L, et al. Omega-3 fatty acids in obesity and metabolic syndrome: a mechanistic update. J Nutr Biochem. 2018;58:1-16. doi:10.1016/j.jnutbio.2018.02.012
46. Kalupahana NS, Claycombe K, Newman SJ, et al. Eicosapentaenoic acid prevents and reverses insulin resistance in high-fat diet-induced obese mice via modulation of adipose tissue inflammation. J Nutr. 2010;140(11):1915-1922. doi:10.3945/jn.110.125732
47. Sze MA, Schloss PD. Looking for a Signal in the Noise: Revisiting Obesity and the Microbiome. mBio. 2016;7(4):10.1128/mbio.01018-16. doi:10.1128/mbio.01018-16
48. Peters BA, Shapiro JA, Church TR, et al. A taxonomic signature of obesity in a large study of American adults. Sci Rep. 2018;8(1):9749. doi:10.1038/s41598-018-28126-1
49. Pesoa SA, Portela N, Fernández E, Elbarcha O, Gotteland M, Magne F. Comparison of Argentinean microbiota with other geographical populations reveals different taxonomic and functional signatures associated with obesity. Sci Rep. 2021;11(1):7762. doi:10.1038/s41598-021-87365-x
50. Stanislawski MA, Dabelea D, Lange LA, Wagner BD, Lozupone CA. Gut microbiota phenotypes of obesity. npj Biofilms Microbiomes. 2019;5(1):1-9. doi:10.1038/s41522-019-0091-8
51. Sun H, Chen D, Yang Y, et al. A High-Fish Oil Diet Can Significantly Reshape The Gut Microbiota In Mice. Published online June 7, 2021. doi:10.21203/rs.3.rs-525477/v1
52. Yang X, Zhang X, Yang W, et al. Gut Microbiota in Adipose Tissue Dysfunction Induced Cardiovascular Disease: Role as a Metabolic Organ. Front Endocrinol (Lausanne). 2021;12:749125. doi:10.3389/fendo.2021.749125
53. Paone P, Suriano F, Jian C, et al. Prebiotic oligofructose protects against high-fat diet-induced obesity by changing the gut microbiota, intestinal mucus production, glycosylation and secretion. Gut Microbes. 2022;14(1):2152307. doi:10.1080/19490976.2022.2152307
54. Huang Y, Ying N, Zhao Q, et al. Amelioration of Obesity-Related Disorders in High-Fat Diet-Fed Mice following Fecal Microbiota Transplantation from Inulin-Dosed Mice. Molecules. 2023;28(10):3997. doi:10.3390/molecules28103997
55. Wang J, Lang T, Shen J, Dai J, Tian L, Wang X. Core Gut Bacteria Analysis of Healthy Mice. Frontiers in Microbiology. 2019;10. Accessed February 14, 2024. https://www.frontiersin.org/journals/microbiology/articles/10.3389/fmicb.2019.00887
56. Nigro E, Scudiero O, Monaco ML, et al. New Insight into Adiponectin Role in Obesity and Obesity-Related Diseases. Biomed Res Int. 2014;2014:658913. doi:10.1155/2014/658913
57. Forny-Germano L, De Felice FG, Vieira MN do N. The Role of Leptin and Adiponectin in Obesity-Associated Cognitive Decline and Alzheimer’s Disease. Frontiers in Neuroscience. 2019;12. Accessed January 17, 2023. https://www.frontiersin.org/articles/10.3389/fnins.2018.01027
58. Obradovic M, Sudar-Milovanovic E, Soskic S, et al. Leptin and Obesity: Role and Clinical Implication. Front Endocrinol (Lausanne). 2021;12:585887. doi:10.3389/fendo.2021.585887
59. Liu W, Zhou X, Li Y, et al. Serum leptin, resistin, and adiponectin levels in obese and non-obese patients with newly diagnosed type 2 diabetes mellitus: A population-based study. Medicine. 2020;99(6):e19052. doi:10.1097/MD.0000000000019052
60. Gariballa S, Alkaabi J, Yasin J, Al Essa A. Total adiponectin in overweight and obese subjects and its response to visceral fat loss. BMC Endocrine Disorders. 2019;19(1):55. doi:10.1186/s12902-019-0386-z
61. Achari AE, Jain SK. Adiponectin, a Therapeutic Target for Obesity, Diabetes, and Endothelial Dysfunction. Int J Mol Sci. 2017;18(6):1321. doi:10.3390/ijms18061321
62. Fabiani H, Mudjihartini N, Lestari W. Dietary Omega-6 to Omega-3 Fatty Acids Ratio is Correlated with High Molecular Weight Adiponectin Level in Indonesian Office Workers. International Journal of Nutrition, Pharmacology, Neurological Diseases. 2021;11(1):64. doi:10.4103/ijnpnd.ijnpnd_89_20
63. Yanai H, Yoshida H. Beneficial Effects of Adiponectin on Glucose and Lipid Metabolism and Atherosclerotic Progression: Mechanisms and Perspectives. Int J Mol Sci. 2019;20(5):1190. doi:10.3390/ijms20051190
64. Pischon T, Girman CJ, Hotamisligil GS, Rifai N, Hu FB, Rimm EB. Plasma Adiponectin Levels and Risk of Myocardial Infarction in Men. JAMA. 2004;291(14): 1730-1737. doi:10.1001/jama.291.14.1730
65. Zocchi M, Della Porta M, Lombardoni F, et al. A Potential Interplay between HDLs and Adiponectin in Promoting Endothelial Dysfunction in Obesity. Biomedicines. 2022;10(6):1344. doi:10.3390/biomedicines10061344
66. Feuerer M, Herrero L, Cipolletta D, et al. Fat Treg cells: a liaison between the immune and metabolic systems. Nat Med. 2009;15(8):930-939. doi:10.1038/nm.2002
67. Cipolletta D, Cohen P, Spiegelman BM, Benoist C, Mathis D. Appearance and disappearance of the mRNA signature characteristic of Treg cells in visceral adipose tissue: Age, diet, and PPARγ effects. Proc Natl Acad Sci U S A. 2015;112(2):482-487. doi:10.1073/pnas.1423486112
68. Lian M, Luo W, Sui Y, Li Z, Hua J. Dietary n-3 PUFA Protects Mice from Con A Induced Liver Injury by Modulating Regulatory T Cells and PPAR-γ Expression. PLoS One. 2015;10(7):e0132741. doi:10.1371/journal.pone.0132741
69. Woodworth HL, McCaskey SJ, Duriancik DM, et al. Dietary fish oil alters T lymphocyte cell populations and exacerbates disease in a mouse model of inflammatory colitis. Cancer Res. 2010;70(20):7960-7969. doi:10.1158/0008-5472.CAN-10-1396
70. Onodera T, Fukuhara A, Jang MH, et al. Adipose tissue macrophages induce PPARγ-high FOXP3+ regulatory T cells. Scientific Reports. 2015;5. doi:10.1038/srep16801
71. Onodera T, Fukuhara A, Shin J, Hayakawa T, Otsuki M, Shimomura I. Eicosapentaenoic acid and 5-HEPE enhance macrophage-mediated Treg induction in mice. Sci Rep. 2017;7(1):4560. doi:10.1038/s41598-017-04474-2
72. Han SC, Koo DH, Kang NJ, et al. Docosahexaenoic Acid Alleviates Atopic Dermatitis by Generating Tregs and IL-10/TGF-β-Modified Macrophages via a TGF-β-Dependent Mechanism. J Invest Dermatol. 2015;135(6):1556-1564. doi:10.1038/jid.2014.488
73. Fooks AN, D’Cruz LM. T Regulatory Cells in the Visceral Adipose Tissues. Immunometabolism. 2021;4(1):e220002. doi:10.20900/immunometab20220002
74. Tan J, Taitz J, Sun SM, Langford L, Ni D, Macia L. Your Regulatory T Cells Are What You Eat: How Diet and Gut Microbiota Affect Regulatory T Cell Development. Front Nutr. 2022;9:878382. doi:10.3389/fnut.2022.878382
75. Rajbhandari P, Arneson D, Hart SK, et al. Single cell analysis reveals immune cell-adipocyte crosstalk regulating the transcription of thermogenic adipocytes. Elife. 2019;8:e49501. doi:10.7554/eLife.49501
76. Beppu LY, Mooli RGR, Qu X, et al. Tregs facilitate obesity and insulin resistance via a Blimp-1/IL-10 axis. JCI Insight. 2021;6(3):e140644. doi:10.1172/jci.insight.140644
77. Atzeni A, Galié S, Muralidharan J, et al. Gut Microbiota Profile and Changes in Body Weight in Elderly Subjects with Overweight/Obesity and Metabolic Syndrome. Microorganisms. 2021;9(2):346. doi:10.3390/microorganisms9020346
78. Wei X, Tao J, Xiao S, et al. Xiexin Tang improves the symptom of type 2 diabetic rats by modulation of the gut microbiota. Sci Rep. 2018;8(1):3685. doi:10.1038/s41598-018-22094-2
79. Lan Y, Ning K, Ma Y, et al. High-Density Lipoprotein Cholesterol as a Potential Medium between Depletion of Lachnospiraceae Genera and Hypertension under a High-Calorie Diet. Microbiol Spectr. 2022;10(6):e02349-22. doi:10.1128/spectrum.02349-22
80. Ofosu FK, Elahi F, Daliri EBM, et al. Fermented sorghum improves type 2 diabetes remission by modulating gut microbiota and their related metabolites in high fat diet-streptozotocin induced diabetic mice. Journal of Functional Foods. 2023;107:105666. doi:10.1016/j.jff.2023.105666
81. Do MH, Lee E, Oh MJ, Kim Y, Park HY. High-Glucose or -Fructose Diet Cause Changes of the Gut Microbiota and Metabolic Disorders in Mice without Body Weight Change. Nutrients. 2018;10(6):761. doi:10.3390/nu10060761
82. Tsai CY, Lu HC, Chou YH, et al. Gut Microbial Signatures for Glycemic Responses of GLP-1 Receptor Agonists in Type 2 Diabetic Patients: A Pilot Study. Front Endocrinol (Lausanne). 2022;12:814770. doi:10.3389/fendo.2021.814770
2. Schwartz MW, Seeley RJ, Zeltser LM, et al. Obesity Pathogenesis: An Endocrine Society Scientific Statement. Endocr Rev. 2017;38(4):267-296. doi:10.1210/er.2017-00111
3. Xu H, Barnes GT, Yang Q, et al. Chronic inflammation in fat plays a crucial role in the development of obesity-related insulin resistance. J Clin Invest. 2003;112(12):1821-1830. doi:10.1172/JCI200319451
4. Ley RE, Bäckhed F, Turnbaugh P, Lozupone CA, Knight RD, Gordon JI. Obesity alters gut microbial ecology. Proc Natl Acad Sci U S A. 2005;102(31):11070-11075. doi:10.1073/pnas.0504978102
5. Ley RE, Turnbaugh PJ, Klein S, Gordon JI. Human gut microbes associated with obesity. Nature. 2006;444(7122):1022-1023. doi:10.1038/4441022a
6. Le Chatelier E, Nielsen T, Qin J, et al. Richness of human gut microbiome correlates with metabolic markers. Nature. 2013;500(7464):541-546. doi:10.1038/nature12506
7. Busebee B, Ghusn W, Cifuentes L, Acosta A. Obesity: A Review of Pathophysiology and Classification. Mayo Clinic Proceedings. 2023;98(12):1842-1857. doi:10.1016/j.mayocp.2023.05.026
8. González-Muniesa P, Mártinez-González MA, Hu FB, et al. Obesity. Nat Rev Dis Primers. 2017;3(1):17034. doi:10.1038/nrdp.2017.34
9. Castoldi A, Naffah de Souza C, Câmara NOS, Moraes-Vieira PM. The Macrophage Switch in Obesity Development. Front Immunol. 2016;6:637. doi:10.3389/fimmu.2015.00637
10. Zatterale F, Longo M, Naderi J, et al. Chronic Adipose Tissue Inflammation Linking Obesity to Insulin Resistance and Type 2 Diabetes. Front Physiol. 2020;10:1607. doi:10.3389/fphys.2019.01607
11. Nishimura S, Manabe I, Nagasaki M, et al. CD8+ effector T cells contribute to macrophage recruitment and adipose tissue inflammation in obesity. Nat Med. 2009;15(8):914-920. doi:10.1038/nm.1964
12. Weisberg SP, McCann D, Desai M, Rosenbaum M, Leibel RL, Ferrante AW. Obesity is associated with macrophage accumulation in adipose tissue. J Clin Invest. 2003;112(12):1796-1808. doi:10.1172/JCI200319246
13. Bradley D, Deng T, Shantaram D, Hsueh WA. Orchestration of the Adipose Tissue Immune Landscape by Adipocytes. Published online February 12, 2024. doi:10.1146/annurev-physiol-042222-024353
14. Chait A, den Hartigh LJ. Adipose tissue distribution, inflammation and its metabolic consequences, including diabetes and cardiovascular disease. Front Cardiovasc Med. 2020;7:22. doi:10.3389/fcvm.2020.00022
15. Turnbaugh PJ, Ley RE, Mahowald MA, Magrini V, Mardis ER, Gordon JI. An obesity-associated gut microbiome with increased capacity for energy harvest. Nature. 2006;444(7122):1027-1031. doi:10.1038/nature05414
16. Cani PD, Amar J, Iglesias MA, et al. Metabolic endotoxemia initiates obesity and insulin resistance. Diabetes. 2007;56(7):1761-1772. doi:10.2337/db06-1491
17. Caesar R, Tremaroli V, Kovatcheva-Datchary P, Cani PD, Bäckhed F. Crosstalk between Gut Microbiota and Dietary Lipids Aggravates WAT Inflammation through TLR Signaling. Cell Metab. 2015;22(4):658-668. doi:10.1016/j.cmet.2015.07.026
18. Haeusler RA, Camastra S, Nannipieri M, et al. Increased Bile Acid Synthesis and Impaired Bile Acid Transport in Human Obesity. J Clin Endocrinol Metab. 2016;101(5):1935-1944. doi:10.1210/jc.2015-2583
19. Agans R, Gordon A, Kramer DL, Perez-Burillo S, Rufián-Henares JA, Paliy O. Dietary Fatty Acids Sustain the Growth of the Human Gut Microbiota. Appl Environ Microbiol. 2018;84(21):e01525-18. doi:10.1128/AEM.01525-18
20. Daniel H, Gholami AM, Berry D, et al. High-fat diet alters gut microbiota physiology in mice. ISME J. 2014;8(2):295-308. doi:10.1038/ismej.2013.155
21. Schoeler M, Caesar R. Dietary lipids, gut microbiota and lipid metabolism. Rev Endocr Metab Disord. 2019;20(4):461-472. doi:10.1007/s11154-019-09512-0
22. Xu AA, Kennedy LK, Hoffman K, et al. Dietary Fatty Acid Intake and the Colonic Gut Microbiota in Humans. Nutrients. 2022;14(13):2722. doi:10.3390/nu14132722
23. López-Millán E, Gutiérrez-Uribe JA, Antunes-Ricardo M. Omega-3 supplementation: Impact on low chronic inflammation associated with obesity. Trends in Food Science & Technology. 2025;155:104799. doi:10.1016/j.tifs.2024.104799
24. Menni C, Jackson MA, Pallister T, Steves CJ, Spector TD, Valdes AM. Gut microbiome diversity and high-fibre intake are related to lower long-term weight gain. Int J Obes. 2017;41(7):1099-1105. doi:10.1038/ijo.2017.66
25. Cao W, Liu F, Li RW, et al. Docosahexaenoic acid-rich fish oil prevented insulin resistance by modulating gut microbiome and promoting colonic peptide YY expression in diet-induced obesity mice. Food Science and Human Wellness. 2022;11(1):177-188. doi:10.1016/j.fshw.2021.07.018
26. Monk JM, Liddle DM, Hutchinson AL, et al. Fish oil supplementation to a high-fat diet improves both intestinal health and the systemic obese phenotype. The Journal of Nutritional Biochemistry. 2019;72:108216. doi:10.1016/j.jnutbio.2019.07.007
27. Watson H, Mitra S, Croden FC, et al. A randomised trial of the effect of omega-3 polyunsaturated fatty acid supplements on the human intestinal microbiota. Gut. 2018;67(11):1974-1983. doi:10.1136/gutjnl-2017-314968
28. Portela ND, Galván C, Sanmarco LM, et al. Omega-3-Supplemented Fat Diet Drives Immune Metabolic Response in Visceral Adipose Tissue by Modulating Gut Microbiota in a Mouse Model of Obesity. Nutrients. 2023;15(6):1404. doi:10.3390/nu15061404
29. Ellacott KLJ, Morton GJ, Woods SC, Tso P, Schwartz MW. Assessment of feeding behavior in laboratory mice. Cell Metab. 2010;12(1):10-17. doi:10.1016/j.cmet.2010.06.001
30. Callahan BJ, McMurdie PJ, Rosen MJ, Han AW, Johnson AJA, Holmes SP. DADA2: High resolution sample inference from Illumina amplicon data. Nat Methods. 2016;13(7):581-583. doi:10.1038/nmeth.3869
31. McMurdie PJ, Holmes S. phyloseq: An R Package for Reproducible Interactive Analysis and Graphics of Microbiome Census Data. PLOS ONE. 2013;8(4):e61217. doi:10.1371/journal.pone.0061217
32. Lahti L, Shetty S. microbiome R package. Published online 2017. doi:10.18129/B9.bioc.microbiome
33. R Core Team. R Core Team (2022). R: A language and environment for statistical computing. Published online 2021. https://www.R-project.org/.
34. Quast C, Pruesse E, Yilmaz P, et al. The SILVA ribosomal RNA gene database project: improved data processing and web-based tools. Nucleic Acids Research. 2013;41(D1):D590-D596. doi:10.1093/nar/gks1219
35. Cao Y, Dong Q, Wang D, Zhang P, Liu Y, Niu C. microbiomeMarker: an R/Bioconductor package for microbiome marker identification and visualization. Bioinformatics. 2022;38(16):4027-4029. doi:10.1093/bioinformatics/btac438
36. Curat CA, Miranville A, Sengenès C, et al. From Blood Monocytes to Adipose Tissue-Resident Macrophages: Induction of Diapedesis by Human Mature Adipocytes. Diabetes. 2004;53(5):1285-1292. doi:10.2337/diabetes.53.5.1285
37. Wei T SV. R package “corrplot”: Visualization of a Correlation Matrix. Published online 2021. https://github.com/taiyun/corrplot.
38. Wickham H. ggplot2: Elegant Graphics for Data Analysis. 2016. Accessed January 25, 2023. https://link.springer.com/book/10.1007/978-3-319-24277-4
39. Kassambara A. ggpubr: “ggplot2” Based Publication Ready Plots. R package version 0.5.0. Published online 2022. https://CRAN.R-project.org/package=ggpubr
40. Dixon P. VEGAN, A Package of R Functions for Community Ecology. Journal of Vegetation Science. 2003;14(6):927-930.
41. Rosseel Y, Jorgensen TD, Wilde LD, et al. lavaan: Latent Variable Analysis. Published online December 20, 2023. Accessed March 7, 2024. https://cran.r-project.org/web/packages/lavaan/index.html
42. Shen W, Wolf PG, Carbonero F, et al. Intestinal and systemic inflammatory responses are positively associated with sulfidogenic bacteria abundance in high-fat-fed male C57BL/6J mice. J Nutr. 2014;144(8):1181-1187. doi:10.3945/jn.114.194332
43. Hildebrandt MA, Hoffmann C, Sherrill-Mix SA, et al. High-fat diet determines the composition of the murine gut microbiome independently of obesity. Gastroenterology. 2009;137(5):1716-1724.e1-2. doi:10.1053/j.gastro.2009.08.042
44. LeMieux MJ, Kalupahana NS, Scoggin S, Moustaid-Moussa N. Eicosapentaenoic acid reduces adipocyte hypertrophy and inflammation in diet-induced obese mice in an adiposity-independent manner. J Nutr. 2015;145(3):411-417. doi:10.3945/jn.114.202952
45. Albracht-Schulte K, Kalupahana NS, Ramalingam L, et al. Omega-3 fatty acids in obesity and metabolic syndrome: a mechanistic update. J Nutr Biochem. 2018;58:1-16. doi:10.1016/j.jnutbio.2018.02.012
46. Kalupahana NS, Claycombe K, Newman SJ, et al. Eicosapentaenoic acid prevents and reverses insulin resistance in high-fat diet-induced obese mice via modulation of adipose tissue inflammation. J Nutr. 2010;140(11):1915-1922. doi:10.3945/jn.110.125732
47. Sze MA, Schloss PD. Looking for a Signal in the Noise: Revisiting Obesity and the Microbiome. mBio. 2016;7(4):10.1128/mbio.01018-16. doi:10.1128/mbio.01018-16
48. Peters BA, Shapiro JA, Church TR, et al. A taxonomic signature of obesity in a large study of American adults. Sci Rep. 2018;8(1):9749. doi:10.1038/s41598-018-28126-1
49. Pesoa SA, Portela N, Fernández E, Elbarcha O, Gotteland M, Magne F. Comparison of Argentinean microbiota with other geographical populations reveals different taxonomic and functional signatures associated with obesity. Sci Rep. 2021;11(1):7762. doi:10.1038/s41598-021-87365-x
50. Stanislawski MA, Dabelea D, Lange LA, Wagner BD, Lozupone CA. Gut microbiota phenotypes of obesity. npj Biofilms Microbiomes. 2019;5(1):1-9. doi:10.1038/s41522-019-0091-8
51. Sun H, Chen D, Yang Y, et al. A High-Fish Oil Diet Can Significantly Reshape The Gut Microbiota In Mice. Published online June 7, 2021. doi:10.21203/rs.3.rs-525477/v1
52. Yang X, Zhang X, Yang W, et al. Gut Microbiota in Adipose Tissue Dysfunction Induced Cardiovascular Disease: Role as a Metabolic Organ. Front Endocrinol (Lausanne). 2021;12:749125. doi:10.3389/fendo.2021.749125
53. Paone P, Suriano F, Jian C, et al. Prebiotic oligofructose protects against high-fat diet-induced obesity by changing the gut microbiota, intestinal mucus production, glycosylation and secretion. Gut Microbes. 2022;14(1):2152307. doi:10.1080/19490976.2022.2152307
54. Huang Y, Ying N, Zhao Q, et al. Amelioration of Obesity-Related Disorders in High-Fat Diet-Fed Mice following Fecal Microbiota Transplantation from Inulin-Dosed Mice. Molecules. 2023;28(10):3997. doi:10.3390/molecules28103997
55. Wang J, Lang T, Shen J, Dai J, Tian L, Wang X. Core Gut Bacteria Analysis of Healthy Mice. Frontiers in Microbiology. 2019;10. Accessed February 14, 2024. https://www.frontiersin.org/journals/microbiology/articles/10.3389/fmicb.2019.00887
56. Nigro E, Scudiero O, Monaco ML, et al. New Insight into Adiponectin Role in Obesity and Obesity-Related Diseases. Biomed Res Int. 2014;2014:658913. doi:10.1155/2014/658913
57. Forny-Germano L, De Felice FG, Vieira MN do N. The Role of Leptin and Adiponectin in Obesity-Associated Cognitive Decline and Alzheimer’s Disease. Frontiers in Neuroscience. 2019;12. Accessed January 17, 2023. https://www.frontiersin.org/articles/10.3389/fnins.2018.01027
58. Obradovic M, Sudar-Milovanovic E, Soskic S, et al. Leptin and Obesity: Role and Clinical Implication. Front Endocrinol (Lausanne). 2021;12:585887. doi:10.3389/fendo.2021.585887
59. Liu W, Zhou X, Li Y, et al. Serum leptin, resistin, and adiponectin levels in obese and non-obese patients with newly diagnosed type 2 diabetes mellitus: A population-based study. Medicine. 2020;99(6):e19052. doi:10.1097/MD.0000000000019052
60. Gariballa S, Alkaabi J, Yasin J, Al Essa A. Total adiponectin in overweight and obese subjects and its response to visceral fat loss. BMC Endocrine Disorders. 2019;19(1):55. doi:10.1186/s12902-019-0386-z
61. Achari AE, Jain SK. Adiponectin, a Therapeutic Target for Obesity, Diabetes, and Endothelial Dysfunction. Int J Mol Sci. 2017;18(6):1321. doi:10.3390/ijms18061321
62. Fabiani H, Mudjihartini N, Lestari W. Dietary Omega-6 to Omega-3 Fatty Acids Ratio is Correlated with High Molecular Weight Adiponectin Level in Indonesian Office Workers. International Journal of Nutrition, Pharmacology, Neurological Diseases. 2021;11(1):64. doi:10.4103/ijnpnd.ijnpnd_89_20
63. Yanai H, Yoshida H. Beneficial Effects of Adiponectin on Glucose and Lipid Metabolism and Atherosclerotic Progression: Mechanisms and Perspectives. Int J Mol Sci. 2019;20(5):1190. doi:10.3390/ijms20051190
64. Pischon T, Girman CJ, Hotamisligil GS, Rifai N, Hu FB, Rimm EB. Plasma Adiponectin Levels and Risk of Myocardial Infarction in Men. JAMA. 2004;291(14): 1730-1737. doi:10.1001/jama.291.14.1730
65. Zocchi M, Della Porta M, Lombardoni F, et al. A Potential Interplay between HDLs and Adiponectin in Promoting Endothelial Dysfunction in Obesity. Biomedicines. 2022;10(6):1344. doi:10.3390/biomedicines10061344
66. Feuerer M, Herrero L, Cipolletta D, et al. Fat Treg cells: a liaison between the immune and metabolic systems. Nat Med. 2009;15(8):930-939. doi:10.1038/nm.2002
67. Cipolletta D, Cohen P, Spiegelman BM, Benoist C, Mathis D. Appearance and disappearance of the mRNA signature characteristic of Treg cells in visceral adipose tissue: Age, diet, and PPARγ effects. Proc Natl Acad Sci U S A. 2015;112(2):482-487. doi:10.1073/pnas.1423486112
68. Lian M, Luo W, Sui Y, Li Z, Hua J. Dietary n-3 PUFA Protects Mice from Con A Induced Liver Injury by Modulating Regulatory T Cells and PPAR-γ Expression. PLoS One. 2015;10(7):e0132741. doi:10.1371/journal.pone.0132741
69. Woodworth HL, McCaskey SJ, Duriancik DM, et al. Dietary fish oil alters T lymphocyte cell populations and exacerbates disease in a mouse model of inflammatory colitis. Cancer Res. 2010;70(20):7960-7969. doi:10.1158/0008-5472.CAN-10-1396
70. Onodera T, Fukuhara A, Jang MH, et al. Adipose tissue macrophages induce PPARγ-high FOXP3+ regulatory T cells. Scientific Reports. 2015;5. doi:10.1038/srep16801
71. Onodera T, Fukuhara A, Shin J, Hayakawa T, Otsuki M, Shimomura I. Eicosapentaenoic acid and 5-HEPE enhance macrophage-mediated Treg induction in mice. Sci Rep. 2017;7(1):4560. doi:10.1038/s41598-017-04474-2
72. Han SC, Koo DH, Kang NJ, et al. Docosahexaenoic Acid Alleviates Atopic Dermatitis by Generating Tregs and IL-10/TGF-β-Modified Macrophages via a TGF-β-Dependent Mechanism. J Invest Dermatol. 2015;135(6):1556-1564. doi:10.1038/jid.2014.488
73. Fooks AN, D’Cruz LM. T Regulatory Cells in the Visceral Adipose Tissues. Immunometabolism. 2021;4(1):e220002. doi:10.20900/immunometab20220002
74. Tan J, Taitz J, Sun SM, Langford L, Ni D, Macia L. Your Regulatory T Cells Are What You Eat: How Diet and Gut Microbiota Affect Regulatory T Cell Development. Front Nutr. 2022;9:878382. doi:10.3389/fnut.2022.878382
75. Rajbhandari P, Arneson D, Hart SK, et al. Single cell analysis reveals immune cell-adipocyte crosstalk regulating the transcription of thermogenic adipocytes. Elife. 2019;8:e49501. doi:10.7554/eLife.49501
76. Beppu LY, Mooli RGR, Qu X, et al. Tregs facilitate obesity and insulin resistance via a Blimp-1/IL-10 axis. JCI Insight. 2021;6(3):e140644. doi:10.1172/jci.insight.140644
77. Atzeni A, Galié S, Muralidharan J, et al. Gut Microbiota Profile and Changes in Body Weight in Elderly Subjects with Overweight/Obesity and Metabolic Syndrome. Microorganisms. 2021;9(2):346. doi:10.3390/microorganisms9020346
78. Wei X, Tao J, Xiao S, et al. Xiexin Tang improves the symptom of type 2 diabetic rats by modulation of the gut microbiota. Sci Rep. 2018;8(1):3685. doi:10.1038/s41598-018-22094-2
79. Lan Y, Ning K, Ma Y, et al. High-Density Lipoprotein Cholesterol as a Potential Medium between Depletion of Lachnospiraceae Genera and Hypertension under a High-Calorie Diet. Microbiol Spectr. 2022;10(6):e02349-22. doi:10.1128/spectrum.02349-22
80. Ofosu FK, Elahi F, Daliri EBM, et al. Fermented sorghum improves type 2 diabetes remission by modulating gut microbiota and their related metabolites in high fat diet-streptozotocin induced diabetic mice. Journal of Functional Foods. 2023;107:105666. doi:10.1016/j.jff.2023.105666
81. Do MH, Lee E, Oh MJ, Kim Y, Park HY. High-Glucose or -Fructose Diet Cause Changes of the Gut Microbiota and Metabolic Disorders in Mice without Body Weight Change. Nutrients. 2018;10(6):761. doi:10.3390/nu10060761
82. Tsai CY, Lu HC, Chou YH, et al. Gut Microbial Signatures for Glycemic Responses of GLP-1 Receptor Agonists in Type 2 Diabetic Patients: A Pilot Study. Front Endocrinol (Lausanne). 2022;12:814770. doi:10.3389/fendo.2021.814770