Digestive Diseases and Sciences

, Volume 62, Issue 2, pp 396–406 | Cite as

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

  • Mitsuaki Ishioka
  • Kouichi Miura
  • Shinichiro Minami
  • Yoichiro Shimura
  • Hirohide Ohnishi
Original Article



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.


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


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.


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.


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


Nonalcoholic fatty liver disease Nonalcoholic steatohepatitis Gut microbiota IL-17 Gut–liver axis 



Alanine transaminase


Choline-deficient amino acid-defined






Hematoxylin and eosin






Nonalcoholic fatty liver disease


Nonalcoholic steatohepatitis


Normal chow


Short-chain fatty acids


Toll-like receptor



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.


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

Compliance with ethical standards

Conflict of interest

All authors have no conflicts to disclose.

Supplementary material

10620_2016_4393_MOESM1_ESM.tiff (2.9 mb)
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)
10620_2016_4393_MOESM2_ESM.tiff (2.9 mb)
Supplementary material 2 (TIFF 2927 kb)
10620_2016_4393_MOESM3_ESM.tiff (2.9 mb)
Supplementary material 3 (TIFF 2927 kb)
10620_2016_4393_MOESM4_ESM.tiff (2.9 mb)
Supplementary material 4 (TIFF 2927 kb)


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Copyright information

© Springer Science+Business Media New York 2016

Authors and Affiliations

  • Mitsuaki Ishioka
    • 1
  • Kouichi Miura
    • 1
  • Shinichiro Minami
    • 1
  • Yoichiro Shimura
    • 2
  • Hirohide Ohnishi
    • 1
  1. 1.Department of GastroenterologyAkita University Graduate School of MedicineAkita-shiJapan
  2. 2.Department of Biotechnology, Faculty of Bioresource SciencesAkita Prefectural UniversityAkita-shiJapan

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