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Altered Gut Microbiota Composition and Immune Response in Experimental Steatohepatitis Mouse Models

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Abstract

Background

Although several types of diet have been used in experimental steatohepatitis models, comparison of gut microbiota and immunological alterations in the gut among diets has not yet been performed.

Aim

We attempted to clarify the difference in the gut environment between mice administrated several experimental diets.

Methods

Male wild-type mice were fed a high-fat (HF) diet, a choline-deficient amino acid-defined (CDAA) diet, and a methionine-choline-deficient (MCD) diet for 8 weeks. We compared the severity of steatohepatitis, the composition of gut microbiota, and the intestinal expression of interleukin (IL)-17, an immune modulator.

Results

Steatohepatitis was most severe in the mice fed the CDAA diet, followed by the MCD diet, and the HF diet. Analysis of gut microbiota showed that the composition of the Firmicutes phylum differed markedly at order level between the mice fed the CDAA and HF diet. The CDAA diet increased the abundance of Clostridiales, while the HF diet increased that of lactate-producing bacteria. In addition, the CDAA diet decreased the abundance of lactate-producing bacteria and antiinflammatory bacterium Parabacteroides goldsteinii in the phylum Bacteroidetes. In CDAA-fed mice, IL-17 levels were increased in ileum as well as portal vein. In addition, the CDAA diet also elevated hepatic expression of chemokines, downstream targets of IL-17.

Conclusions

The composition of gut microbiota and IL-17 expression varied considerably between mice administrated different experimental diets to induce steatohepatitis.

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Abbreviations

ALT:

Alanine transaminase

CDAA:

Choline-deficient amino acid-defined

F/B :

Firmicutes/Bacteroidetes

HF:

High-fat

H&E:

Hematoxylin and eosin

IL:

Interleukin

MCD:

Methionine-choline-deficient

NAFLD:

Nonalcoholic fatty liver disease

NASH:

Nonalcoholic steatohepatitis

NC:

Normal chow

SCFAs:

Short-chain fatty acids

TLR:

Toll-like receptor

References

  1. Okanoue T, Umemura A, Yasui K, et al. Nonalcoholic fatty liver disease and nonalcoholic steatohepatitis in Japan. J Gastroenterol Hepatol. 2011;1:153–162.

    Article  Google Scholar 

  2. Mouzaki M, Comelli EM, Arendt BM, et al. Intestinal microbiota in patients with nonalcoholic fatty liver disease. Hepatology. 2013;58:120–127.

    Article  CAS  PubMed  Google Scholar 

  3. Miura K, Ohnishi H. Role of gut microbiota and toll-like receptors in nonalcoholic fatty liver disease. World J Gastroenterol. 2014;20:7381–7391.

    Article  PubMed  PubMed Central  Google Scholar 

  4. Bäckhed F, Ley RE, Sonnenburg JL, et al. Host-bacterial mutualism in the human intestine. Science. 2005;307:1915–1920.

    Article  PubMed  Google Scholar 

  5. Ley RE, Turnbaugh PJ, Klein S, et al. Microbial ecology: human gut microbes associated with obesity. Nature. 2006;444:1022–1023.

    Article  CAS  PubMed  Google Scholar 

  6. Mathur R, Barlow GM. Obesity and the microbiome. Expert Rev Gastroenterol Hepatol. 2015;9:1087–1099.

    Article  CAS  PubMed  Google Scholar 

  7. Damms-Machado A, Mitra S, Schollenberger AE, et al. Effects of surgical and dietary weight loss therapy for obesity on gut microbiota composition and nutrient absorption. Biomed Res Int. 2015;2015:806248.

    Article  PubMed  PubMed Central  Google Scholar 

  8. Huang Y, Fan XG, Wang ZM, et al. Identification of helicobacter species in human liver samples from patients with primary hepatocellular carcinoma. J Clin Pathol. 2004;57:1273–1277.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  9. Ferolla SM, Armiliato GN, Couto CA, et al. The role of intestinal bacteria overgrowth in obesity-related nonalcoholic fatty liver disease. Nutrients. 2014;6:5583–5599.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  10. Zhu L, Baker SS, Gill C, et al. Characterization of gut microbiomes in nonalcoholic steatohepatitis (NASH) patients: a connection between endogenous alcohol and NASH. Hepatology. 2013;57:601–609.

    Article  CAS  PubMed  Google Scholar 

  11. Okubo H, Sakoda H, Kushiyama A, et al. Lactobacillus casei strain Shirota protects against nonalcoholic steatohepatitis development in a rodent model. Am J Physiol Gastrointest Liver Physiol. 2013;305:G911–G918.

    Article  CAS  PubMed  Google Scholar 

  12. Naito E, Yoshida Y, Makino K, et al. Beneficial effect of oral administration of Lactobacillus casei strain Shirota on insulin resistance in diet-induced obesity mice. J Appl Microbiol. 2011;110:650–657.

    Article  CAS  PubMed  Google Scholar 

  13. Ivanov II, Frutos Rde L, Manel N, et al. Specific microbiota direct the differentiation of IL-17-producing T-helper cells in the mucosa of the small intestine. Cell Host Microbe. 2008;4:337–349.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  14. Tang Y, Bian Z, Zhao L, et al. Interleukin-17 exacerbates hepatic steatosis and inflammation in nonalcoholic fatty liver disease. Clin Exp Immunol. 2011;166:281–290.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  15. Ibrahim SH, Hirsova P, Malhi H, et al. Animal models of nonalcoholic steatohepatitis: Eat, delete, and inflame. Dig Dis Sci. 2016;61:1325–1336.

    Article  PubMed  Google Scholar 

  16. Yamaguchi K, Yang L, McCall S, et al. Inhibiting triglyceride synthesis improves hepatic steatosis but exacerbates liver damage and fibrosis in obese mice with nonalcoholic steatohepatitis. Hepatology. 2007;45:1366–1374.

    Article  CAS  PubMed  Google Scholar 

  17. Hebbard L, George J. Animal models of nonalcoholic fatty liver disease. Nat Rev Gastroenterol Hepatol. 2011;8:35–44.

    Article  PubMed  Google Scholar 

  18. Ito M, Suzuki J, Tsujioka S, et al. Longitudinal analysis of murine steatohepatitis model induced by chronic exposure to high-fat diet. Hepatol Res. 2007;37:50–57.

    Article  CAS  PubMed  Google Scholar 

  19. Miura K, Kodama Y, Inokuchi S, et al. Toll-like receptor 9 promotes steatohepatitis by induction of interleukin-1beta in mice. Gastroenterology. 2010;139:e7.

    Article  Google Scholar 

  20. Miura K, Ishioka M, Minami S, et al. Toll-like receptor 4 on macrophage promotes the development of steatohepatitis-related hepatocellular carcinoma in mice. J Biol Chem. 2016;27:11504–11517.

    Article  Google Scholar 

  21. Brunt EM, Kleiner DE, Wilson LA, et al. NASH clinical research network (CRN). Nonalcoholic fatty liver disease (NAFLD) activity score and the histopathologic diagnosis in NAFLD: distinct clinicopathologic meanings. Hepatology. 2011;53:810–820.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  22. Loman NJ, Misra RV, Dallman TJ, et al. Performance comparison of benchtop high-throughput sequencing platforms. Nat Biotechnol. 2012;30:434–439.

    Article  CAS  PubMed  Google Scholar 

  23. Wright ES, Yilmaz LS, Noguera DR. DECIPHER, a search-based approach to chimera identification for 16S rRNA sequences. Appl Environ Microbiol. 2012;78:717–725.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  24. Kitani H, Takenouchi T, Sato M, Yoshioka M, Yamanaka N. A simple and efficient method to isolate macrophages from mixed primary cultures of adult liver cells. J Vis Exp JoVE. 2011;e0000.

  25. Petit E, LaTouf WG, Coppi MV, et al. Involvement of a bacterial microcompartment in the metabolism of fucose and rhamnose by Clostridium phytofermentans. PLoS ONE. 2013;8:e54337.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  26. Collins MD, Lawson PA, Willems A, et al. The phylogeny of the genus Clostridium: proposal of five new genera and eleven new species combinations. Int J Syst Bacteriol. 1994;44:812–826.

    Article  CAS  PubMed  Google Scholar 

  27. Tuck LR, Altenbach K, Ang TF, et al. Insight into Coenzyme A cofactor binding and the mechanism of acyl-transfer in an acylating aldehyde dehydrogenase from Clostridium phytofermentans. Sci Rep. 2016;6:22108.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  28. Chang CJ, Lin CS, Lu CC, et al. Ganoderma lucidum reduces obesity in mice by modulating the composition of the gut microbiota. Nat Commun. 2015;6:7489.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  29. Neyrinck AM, Etxeberria U, Taminiau B, et al. Rhubarb extract prevents hepatic inflammation induced by acute alcohol intake, an effect related to the modulation of the gut microbiota. Mol Nutr Food Res. 2016. doi:10.1002/mnfr.201500899.

  30. Chen Y, Yang F, Lu H, et al. Characterization of fecal microbial communities in patients with liver cirrhosis. Hepatology. 2011;54:562–572.

    Article  PubMed  Google Scholar 

  31. Chinen T, Rudensky AY. The effects of commensal microbiota on immune cell subsets and inflammatory responses. Immunol Rev. 2012;245:45–55.

    Article  CAS  PubMed  Google Scholar 

  32. Lemmers A, Moreno C, Gustot T, et al. The interleukin-17 pathway is involved in human alcoholic liver disease. Hepatology. 2009;49:646–657.

    Article  CAS  PubMed  Google Scholar 

  33. Zhang Y, Chen L, Gao W, et al. IL-17 neutralization significantly ameliorates hepatic granulomatous inflammation and liver damage in Schistosoma japonicum infected mice. Eur J Immunol. 2012;42:1523–1535.

    Article  CAS  PubMed  Google Scholar 

  34. Delzenne NM, Cani PD. Interaction between obesity and the gut microbiota: relevance in nutrition. Annu Rev Nutr. 2011;31:15–31.

    Article  CAS  PubMed  Google Scholar 

  35. Sakamoto M. Reclassification of Bacteroides distasonis, Bacteroides goldsteinii and Bacteroides merdae as Parabacteroides distasonis gen. nov., comb. nov., Parabacteroides goldsteinii comb. nov. and Parabacteroides merdae comb. nov. Int J Syst Evol Microbiol. 2006;56:1599–1605.

    Article  CAS  PubMed  Google Scholar 

  36. Pfalzer AC, Nesbeth PD, Parnell LD, et al. Diet- and genetically-induced obesity differentially affect the fecal microbiome and metabolome in Apc1638N mice. PLoS ONE. 2015;10:e0135758.

    Article  PubMed  PubMed Central  Google Scholar 

  37. Kverka M, Zakostelska Z, Klimesova K, et al. Oral administration of Parabacteroides distasonis antigens attenuates experimental murine colitis through modulation of immunity and microbiota composition. Clin Exp Immunol. 2011;163:250–259.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  38. Murphy EF, Cotter PD, Healy S, et al. Composition and energy harvesting capacity of the gut microbiota: relationship to diet, obesity and time in mouse models. Gut. 2010;59:1635–1642.

    Article  CAS  PubMed  Google Scholar 

  39. Segura-López FK, Güitrón-Cantú A, Torres J. Association between Helicobacter spp. infections and hepatobiliary malignancies: a review. World J Gastroenterol. 2015;21:1414–1423.

    Article  PubMed  PubMed Central  Google Scholar 

  40. Goldstein EJ, Citron DM, Peraino VA, et al. Desulfovibrio desulfuricans bacteremia and review of human Desulfovibrio infections. J Clin Microbiol. 2003;41:2752–2754.

    Article  PubMed  PubMed Central  Google Scholar 

  41. Imajo K, Fujita K, Yoneda M, et al. Hyperresponsivity to low-dose endotoxin during progression to nonalcoholic steatohepatitis is regulated by leptin-mediated signaling. Cell Metab. 2012;16:44–54.

    Article  CAS  PubMed  Google Scholar 

  42. D’Argenio V, Casaburi G, Precone V, Salvatore F. Comparative metagenomic analysis of human gut microbiome composition using two different bioinformatic pipelines. BioMed Res Int. 2014;2014:1–10.

    Article  Google Scholar 

  43. Rolla S, Alchera E, Imarisio C, et al. The balance between IL-17 and IL-22 produced by liver-infiltrating T-helper cells critically controls NASH development in mice. Clin Sci (Lond). 2016;130:193–203.

    Article  CAS  Google Scholar 

  44. Takahashi N, Vanlaere I, de Rycke R, et al. IL-17 produced by Paneth cells drives TNF-induced shock. J Exp Med. 2008;205:1755–1761.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  45. Park SW, Kim M, Brown KM, et al. Paneth cell-derived interleukin-17A causes multiorgan dysfunction after hepatic ischemia and reperfusion injury. Hepatology. 2011;53:1662–1675.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  46. Jin W, Dong C. IL-17 cytokines in immunity and inflammation. Emerg Microbes Infect. 2013;2:e60.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  47. Miura K, Yang L, van Rooijen N, et al. Hepatic recruitment of macrophages promotes nonalcoholic steatohepatitis through CCR2. Am J Physiol Gastrointest Liver Physiol. 2012;302:G1310–G1321.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  48. Gomes AC, Bueno AA, de Souza RG, et al. Gut microbiota, probiotics and diabetes. Nutr J. 2014;13:60.

    Article  PubMed  PubMed Central  Google Scholar 

  49. Haileselassie Y, Johansson MA, Zimmer CL, et al. Lactobacilli regulate Staphylococcus aureus 161:2-induced pro-inflammatory T-cell responses in vitro. PLoS ONE. 2013;8:e77893.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  50. Geem D, Medina-Contreras O, McBride M, et al. Specific microbiota-induced intestinal Th17 differentiation requires MHC class II but not GALT and mesenteric lymph nodes. J Immunol. 2014;193:431–438.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  51. Harley IT, Stankiewicz TE, Giles DA, et al. IL-17 signaling accelerates the progression of nonalcoholic fatty liver disease in mice. Hepatology. 2014;59:1830–1839.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

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Acknowledgments

We thank Daichi Nakagawa for excellent technical assistance and the Biotechnology Center, Faculty of Bioresource Sciences, Akita Prefectural University, for assistance with the GS Junior instrument for pyrosequencing analyses.

Funding

This study was supported by a Grant-in-Aid for Scientific Research from the Japan Society for the Promotion of Science (K.M.).

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Authors and Affiliations

  1. Department of Gastroenterology, Akita University Graduate School of Medicine, 1-1-1 Hondo, Akita-shi, Akita, 010-8543, Japan

    Mitsuaki Ishioka, Kouichi Miura, Shinichiro Minami & Hirohide Ohnishi

  2. Department of Biotechnology, Faculty of Bioresource Sciences, Akita Prefectural University, 241-438 Kaidobata-Nishi, Shimoshinjo-Nakano, Akita-shi, Akita, 010-0195, Japan

    Yoichiro Shimura

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  1. Mitsuaki Ishioka
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Correspondence to Kouichi Miura.

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Supplemental Figure 1

Composition of gut microbiota examined by the GS Junior system. The relative abundance of gut microbiota is shown for HF, CDAA, and MCD diets, compared with the NC group. First, the percentage of the gut microbiota was calculated in the total count reads (NC: 10,829, HF: 13,374, CDAA: 26,009, MCD: 22,601 reads). Red indicates increased abundance, while green indicates decreased abundance, compared with the NC group. The composition of the gut microbiota is shown at phylum, order, genus, and species level. (A) Composition of gut microbiota in phylum Firmicutes. “C” indicates Clostridium. (B) Composition of gut microbiota in phylum Bacteroidetes. “B” and “P” indicate Bacteroides and Parabacteroides, respectively. (C) Composition of gut microbiota in phylum Proteobacteria. “D” and “H” indicate Desulfovibrio and Helicobacter, respectively. (TIFF 2927 kb)

Supplementary material 2 (TIFF 2927 kb)

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Supplementary material 4 (TIFF 2927 kb)

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Ishioka, M., Miura, K., Minami, S. et al. Altered Gut Microbiota Composition and Immune Response in Experimental Steatohepatitis Mouse Models. Dig Dis Sci 62, 396–406 (2017). https://doi.org/10.1007/s10620-016-4393-x

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  • DOI: https://doi.org/10.1007/s10620-016-4393-x

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