Top

Published in:

01-10-2012 | Article

Modulation of gut microbiota by antibiotics improves insulin signalling in high-fat fed mice

Authors: B. M. Carvalho, D. Guadagnini, D. M. L. Tsukumo, A. A. Schenka, P. Latuf-Filho, J. Vassallo, J. C. Dias, L. T. Kubota, J. B. C. Carvalheira, M. J. A. Saad

Published in: Diabetologia | Issue 10/2012

Abstract

Aims/hypothesis

A high-fat dietary intake induces obesity and subclinical inflammation, which play important roles in insulin resistance. Recent studies have suggested that increased concentrations of circulating lipopolysaccharide (LPS), promoted by changes in intestinal permeability, may have a pivotal role in insulin resistance. Thus, we investigated the effect of gut microbiota modulation on insulin resistance and macrophage infiltration.

Methods

Swiss mice were submitted to a high-fat diet with antibiotics or pair-feeding for 8 weeks. Metagenome analyses were performed on DNA samples from mouse faeces. Blood was collected to determine levels of glucose, insulin, LPS, cytokines and acetate. Liver, muscle and adipose tissue proteins were analysed by western blotting. In addition, liver and adipose tissue were analysed, blinded, using histology and immunohistochemistry.

Results

Antibiotic treatment greatly modified the gut microbiota, reducing levels of Bacteroidetes and Firmicutes, overall bacterial count and circulating LPS levels. This modulation reduced levels of fasting glucose, insulin, TNF-α and IL-6; reduced activation of toll-like receptor 4 (TLR4), c-Jun N-terminal kinase (JNK), inhibitor of κ light polypeptide gene enhancer in B cells, kinase β (IKKβ) and phosphorylated IRS-1 Ser307; and consequently improved glucose tolerance and insulin tolerance and action in metabolically active tissues. In addition, there was an increase in portal levels of circulating acetate, which probably contributed to an increase in 5′-AMP-activated protein kinase (AMPK) phosphorylation in mice. We observed a striking reduction in crown-like structures (CLS) and F4/80⁺ macrophage cells in the adipose tissue of antibiotic-treated mice.

Conclusions/interpretation

These results suggest that modulation of gut microbiota in obesity can improve insulin signalling and glucose tolerance by reducing circulating LPS levels and inflammatory signalling. Modulation also appears to increase levels of circulating acetate, which activates AMPK and finally leads to reduced macrophage infiltration.

Available only for authorised users

Kahn SE, Hull RL, Utzschneider KM (2006) Mechanisms linking obesity to insulin resistance and type 2 diabetes. Nature 444:840–846PubMedCrossRef

Wellen KE, Hotamisligil GS (2005) Inflammation, stress, and diabetes. J Clin Invest 115:1111–1119PubMed

Backhed F, Ding H, Wang T et al (2004) The gut microbiota as an environmental factor that regulates fat storage. Proc Natl Acad Sci USA 101:15718–15723PubMedCrossRef

Turnbaugh PJ, Ley RE, Mahowald MA, Magrini V, Mardis ER, Gordon JI (2006) An obesity-associated gut microbiome with increased capacity for energy harvest. Nature 444:1027–1031PubMedCrossRef

Turnbaugh PJ, Backhed F, Fulton L, Gordon JI (2008) Diet-induced obesity is linked to marked but reversible alterations in the mouse distal gut microbiome. Cell Host Microbe 3:213–223PubMedCrossRef

Ley RE, Backhed F, Turnbaugh P, Lozupone CA, Knight RD, Gordon JI (2005) Obesity alters gut microbial ecology. Proc Natl Acad Sci USA 102:11070–11075PubMedCrossRef

Zhang H, DiBaise JK, Zuccolo A et al (2009) Human gut microbiota in obesity and after gastric bypass. Proc Natl Acad Sci USA 106:2365–2370PubMedCrossRef

Schumann A, Nutten S, Donnicola D et al (2005) Neonatal antibiotic treatment alters gastrointestinal tract developmental gene expression and intestinal barrier transcriptome. Physiol Genomics 23:235–245PubMedCrossRef

Membrez M, Blancher F, Jaquet M et al (2008) Gut microbiota modulation with norfloxacin and ampicillin enhances glucose tolerance in mice. FASEB J 22:2416–2426PubMedCrossRef

10.

Carvalho-Filho MA, Ropelle ER, Pauli RJ et al (2009) Aspirin attenuates insulin resistance in muscle of diet-induced obese rats by inhibiting inducible nitric oxide synthase production and S-nitrosylation of IRbeta/IRS-1 and Akt. Diabetologia 52:2425–2434PubMedCrossRef

11.

Schenka AA, Machado CM, Grippo MC et al (2005) Immunophenotypic and ultrastructural validation of a new human glioblastoma cell line. Cell Mol Neurobiol 25:929–941PubMedCrossRef

12.

Cintra DE, Pauli JR, Araujo EP et al (2008) Interleukin-10 is a protective factor against diet-induced insulin resistance in liver. J Hepatol 48:628–637PubMedCrossRef

13.

Schwiertz A, Taras D, Schafer K et al (2010) Microbiota and SCFA in lean and overweight healthy subjects. Obesity 18:190–195PubMedCrossRef

14.

Collado MC, Isolauri E, Laitinen K, Salminen S (2008) Distinct composition of gut microbiota during pregnancy in overweight and normal-weight women. Am J Clin Nutr 88:894–899PubMed

15.

Henao-Mejia J, Elinav E, Jin C et al (2012) Inflammasome-mediated dysbiosis regulates progression of NAFLD and obesity. Nature 482:179–185PubMed

16.

Brun P, Castagliuolo I, Di Leo V et al (2007) Increased intestinal permeability in obese mice: new evidence in the pathogenesis of nonalcoholic steatohepatitis. Am J Physiol Gastrointest Liver Physiol 292:G518–G525PubMedCrossRef

17.

Cani PD, Neyrinck AM, Fava F et al (2007) Selective increases of bifidobacteria in gut microflora improve high-fat-diet-induced diabetes in mice through a mechanism associated with endotoxaemia. Diabetologia 50:2374–2383PubMedCrossRef

18.

Carvalho-Filho MA, Ueno M, Carvalheira JB, Velloso LA, Saad MJ (2006) Targeted disruption of iNOS prevents LPS-induced S-nitrosation of IRbeta/IRS-1 and Akt and insulin resistance in muscle of mice. Am J Physiol Endocrinol Metab 291:E476–E482PubMedCrossRef

19.

Oliveira AG, Carvalho BM, Tobar N et al (2011) Physical exercise reduces circulating lipopolysaccharide and TLR4 activation and improves insulin signaling in tissues of DIO rats. Diabetes 60:784–796PubMedCrossRef

20.

Calisto KL, Carvalho Bde M, Ropelle ER et al (2010) Atorvastatin improves survival in septic rats: effect on tissue inflammatory pathway and on insulin signaling. PLoS One 5:e14232PubMedCrossRef

21.

Tsukumo DM, Carvalho-Filho MA, Carvalheira JB et al (2007) Loss-of-function mutation in Toll-like receptor 4 prevents diet-induced obesity and insulin resistance. Diabetes 56:1986–1998PubMedCrossRef

22.

Shi H, Kokoeva MV, Inouye K, Tzameli I, Yin H, Flier JS (2006) TLR4 links innate immunity and fatty acid-induced insulin resistance. J Clin Invest 116:3015–3025PubMedCrossRef

23.

Pilon G, Charbonneau A, White PJ et al (2010) Endotoxin mediated-iNOS induction causes insulin resistance via ONOO induced tyrosine nitration of IRS-1 in skeletal muscle. PLoS One 5:e15912PubMedCrossRef

24.

Song MJ, Kim KH, Yoon JM, Kim JB (2006) Activation of Toll-like receptor 4 is associated with insulin resistance in adipocytes. Biochem Biophys Res Commun 346:739–745PubMedCrossRef

25.

Nguyen MT, Favelyukis S, Nguyen AK et al (2007) A subpopulation of macrophages infiltrates hypertrophic adipose tissue and is activated by free fatty acids via Toll-like receptors 2 and 4 and JNK-dependent pathways. J Biol Chem 282:35279–35292PubMedCrossRef

26.

Cani PD, Bibiloni R, Knauf C et al (2008) Changes in gut microbiota control metabolic endotoxemia-induced inflammation in high-fat diet-induced obesity and diabetes in mice. Diabetes 57:1470–1481PubMedCrossRef

27.

De Silva A, Bloom SR (2012) Gut hormones and appetite control: a focus on PYY and GLP-1 as therapeutic targets in obesity. Gut Liver 6:10–20PubMedCrossRef

28.

Cani PD, Possemiers S, van de Wiele T et al (2009) Changes in gut microbiota control inflammation in obese mice through a mechanism involving GLP-2-driven improvement of gut permeability. Gut 58:1091–1103PubMedCrossRef

29.

Saad MJ, Araki E, Miralpeix M, Rothenberg PL, White MF, Kahn CR (1992) Regulation of insulin receptor substrate-1 in liver and muscle of animal models of insulin resistance. J Clin Invest 90:1839–1849PubMedCrossRef

30.

Okazaki Y, Tomotake H, Tsujimoto K, Sasaki M, Kato N (2011) Consumption of a resistant protein, sericin, elevates fecal immunoglobulin a, mucins, and cecal organic acids in rats fed a high-fat diet. J Nutr 141:1975–1981PubMedCrossRef

31.

Sakakibara S, Yamauchi T, Oshima Y, Tsukamoto Y, Kadowaki T (2006) Acetic acid activates hepatic AMPK and reduces hyperglycemia in diabetic KK-A(y) mice. Biochem Biophys Res Commun 344:597–604PubMedCrossRef

32.

Kahn BB, Alquier T, Carling D, Hardie DG (2005) AMP-activated protein kinase: ancient energy gauge provides clues to modern understanding of metabolism. Cell Metab 1:15–25PubMedCrossRef

33.

Ishizuka S, Tanaka S, Xu H, Hara H (2004) Fermentable dietary fiber potentiates the localization of immune cells in the rat large intestinal crypts. Exp Biol Med 229:876–884

34.

Shimotoyodome A, Meguro S, Hase T, Tokimitsu I, Sakata T (2000) Short chain fatty acids but not lactate or succinate stimulate mucus release in the rat colon. Comp Biochem Physiol A Mol Integr Physiol 125:525–531PubMedCrossRef

35.

Everard A, Lazarevic V, Derrien M et al (2011) Responses of gut microbiota and glucose and lipid metabolism to prebiotics in genetic obese and diet-induced leptin-resistant mice. Diabetes 60:2775–2786PubMedCrossRef

36.

Ben-Shlomo S, Zvibel I, Shnell M et al (2011) Glucagon-like peptide-1 reduces hepatic lipogenesis via activation of AMP-activated protein kinase. J Hepatol 54:1214–1223PubMedCrossRef

37.

Kang K, Reilly SM, Karabacak V et al (2008) Adipocyte-derived Th2 cytokines and myeloid PPARdelta regulate macrophage polarization and insulin sensitivity. Cell Metab 7:485–495PubMedCrossRef

38.

Odegaard JI, Ricardo-Gonzalez RR, Red Eagle A et al (2008) Alternative M2 activation of Kupffer cells by PPARdelta ameliorates obesity-induced insulin resistance. Cell Metab 7:496–507PubMedCrossRef

39.

Spooner CJ, Sebastian T, Shuman JD et al (2007) C/EBPbeta serine 64, a phosphoacceptor site, has a critical role in LPS-induced IL-6 and MCP-1 transcription. Cytokine 37:119–127PubMedCrossRef

40.

Yamana J, Santos L, Morand E (2009) Enhanced induction of LPS-induced fibroblast MCP-1 by interferon-gamma: involvement of JNK and MAPK phosphatase-1. Cell Immunol 255:26–32PubMedCrossRef

41.

Jiang C, Ting AT, Seed B (1998) PPAR-gamma agonists inhibit production of monocyte inflammatory cytokines. Nature 391:82–86PubMedCrossRef

42.

Ricote M, Li AC, Willson TM, Kelly CJ, Glass CK (1998) The peroxisome proliferator-activated receptor-gamma is a negative regulator of macrophage activation. Nature 391:79–82PubMedCrossRef

43.

Cancello R, Henegar C, Viguerie N et al (2005) Reduction of macrophage infiltration and chemoattractant gene expression changes in white adipose tissue of morbidly obese subjects after surgery-induced weight loss. Diabetes 54:2277–2286PubMedCrossRef

44.

Kováčiková M, Sengenes C, Kováčová Z et al (2011) Dietary intervention-induced weight loss decreases macrophage content in adipose tissue of obese women. Int J Obes 35:91–98CrossRef

Title: Modulation of gut microbiota by antibiotics improves insulin signalling in high-fat fed mice
Authors: B. M. Carvalho
D. Guadagnini
D. M. L. Tsukumo
A. A. Schenka
P. Latuf-Filho
J. Vassallo
J. C. Dias
L. T. Kubota
J. B. C. Carvalheira
M. J. A. Saad
Publication date: 01-10-2012
Publisher: Springer-Verlag
Published in: Diabetologia / Issue 10/2012
Print ISSN: 0012-186X
Electronic ISSN: 1432-0428
DOI: https://doi.org/10.1007/s00125-012-2648-4

ACC 2024 Congress Coverage

Heart Failure Video

At a glance: The ONWARDS insulin icodec trials

Springer Medicine

Modulation of gut microbiota by antibiotics improves insulin signalling in high-fat fed mice

Abstract

Aims/hypothesis

Methods

Results

Conclusions/interpretation

ACC 2024 Congress Coverage

Year in Review: Heart failure and cardiomyopathies

Semaglutide benefits people with type 2 diabetes and obesity-related heart failure

Incentivised to exercise

New intervention for STEMI-related cardiogenic shock

» View more highlights on the ACC 2024 congress hub page

At a glance: The ONWARDS insulin icodec trials

Springer Medicine

Abstract

Aims/hypothesis

Methods

Results

Conclusions/interpretation

Please log in to get access to this content

Other articles of this Issue 10/2012

Prevalence of monogenic diabetes amongst Polish children after a nationwide genetic screening campaign

GATA6 inactivating mutations are associated with heart defects and, inconsistently, with pancreatic agenesis and diabetes

Controversy about the relative efficacy of dipeptidyl peptidase IV inhibitors

Low doses of anti-CD3, ciclosporin A and the vitamin D analogue, TX527, synergise to delay recurrence of autoimmune diabetes in an islet-transplanted NOD mouse model of diabetes

Adipocyte-released insulin-like growth factor-1 is regulated by glucose and fatty acids and controls breast cancer cell growth in vitro

Systemic inflammation in childhood obesity: circulating inflammatory mediators and activated CD14++ monocytes

ACC 2024 Congress Coverage

Year in Review: Heart failure and cardiomyopathies

Semaglutide benefits people with type 2 diabetes and obesity-related heart failure

Incentivised to exercise

New intervention for STEMI-related cardiogenic shock

» View more highlights on the ACC 2024 congress hub page