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Research Article
Longitudinal characterization of diet-induced genetic murine models of non-alcoholic steatohepatitis with metabolic, histological, and transcriptomic hallmarks of human patients
Naomichi Abe, Sayuka Kato, Takuma Tsuchida, Kanami Sugimoto, Ryuta Saito, Lars Verschuren, Robert Kleemann, Kozo Oka
Biology Open 2019 8: bio041251 doi: 10.1242/bio.041251 Published 1 May 2019
Naomichi Abe
1Sohyaku. Innovative Research Division, Mitsubishi Tanabe Pharma Corporation, 2-2-50, Kawagishi, Toda-shi, Saitama 335-8505, Japan
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  • ORCID record for Naomichi Abe
  • For correspondence: Abe.Naomichi@me.mt-pharma.co.jp
Sayuka Kato
1Sohyaku. Innovative Research Division, Mitsubishi Tanabe Pharma Corporation, 2-2-50, Kawagishi, Toda-shi, Saitama 335-8505, Japan
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Takuma Tsuchida
1Sohyaku. Innovative Research Division, Mitsubishi Tanabe Pharma Corporation, 2-2-50, Kawagishi, Toda-shi, Saitama 335-8505, Japan
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Kanami Sugimoto
1Sohyaku. Innovative Research Division, Mitsubishi Tanabe Pharma Corporation, 2-2-50, Kawagishi, Toda-shi, Saitama 335-8505, Japan
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Ryuta Saito
1Sohyaku. Innovative Research Division, Mitsubishi Tanabe Pharma Corporation, 2-2-50, Kawagishi, Toda-shi, Saitama 335-8505, Japan
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Lars Verschuren
2Department of Microbiology and Systems Biology, Netherlands Organization for Applied Scientific Research (TNO), Utrechtseweg 48, 3700 AJ, Zeist, The Netherlands
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Robert Kleemann
3Department of Metabolic Health Research, Netherlands Organization for Applied Scientific Research (TNO), Zernikedreef 9, 2301 CE, Leiden, the Netherlands
4Department of Vascular Surgery, Netherlands Organization for Applied Scientific Research (TNO), Albinusdreef 2, P.O.Box 9600, 2333 ZA Leiden, The Netherlands
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Kozo Oka
1Sohyaku. Innovative Research Division, Mitsubishi Tanabe Pharma Corporation, 2-2-50, Kawagishi, Toda-shi, Saitama 335-8505, Japan
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    Fig. 1.

    Body weight and food intake changes in C57BL/6J, ob/ob and KK-Ay mice under NCD, FFD, or CDAHFD regimes for 30 weeks. Body weight (A–C) and food intake (D–F). Values are mean±s.e.m., n=5. *P<0.05, **P<0.01 versus NCD (Student's t-test, two-tailed).

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    Fig. 2.

    Plasma insulin, plasma ALT levels and hepatic cholesterol levels in C57BL/6J, ob/ob and KK-Ay mice under NCD, FFD, or CDAHFD regimes for 2, 4, 8, 12, 18, 24 and 30 weeks. Plasma insulin levels (A–C), plasma ALT levels (D–F) and hepatic cholesterol levels (G–I). Values are mean±s.e.m., n=3–5 (the exact number of animals are shown in the figure). *P<0.05, **P<0.01 versus NCD (Student's t-test, two-tailed).

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    Fig. 3.

    Histological changes in C57BL/6J, ob/ob and KK-Ay mice under NCD, FFD, or CDAHFD regimes for 2, 4, 8, 12, 18, 24 and 30 weeks. (A) Hematoxylin and Eosin staining of liver sections at 30 weeks. Scale bars: 200 µm. (B) Sirius Red staining of liver sections at 30 weeks. Scale bars: 50 µm. NAFLD activity score (NAS) (C–E) and Sirius Red-positive area (%) (F–H). Values are mean±s.e.m., n=4–5 (the exact number of animals are shown in the figure). *P<0.05, **P<0.01 versus NCD (NAS: Wilcoxon's test, Sirius Red-positive area: Student's t-test, two-tailed).

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    Fig. 4.

    Histological changes of hCLS in ob/ob mice under NCD or FFD regimes. (A,B) F4/80 staining of liver sections at 12 weeks, (C) hCLS numbers (cells/field) at 2, 4, 8, 12, 18, 24 and 30 weeks. Values are mean±s.e.m., n=4–5 (the exact number of animals are shown in the figure). *P<0.05, **P<0.01 versus NCD (Student's t-test, two-tailed). Scale bars: 50 µm.

  • Fig. 5.
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    Fig. 5.

    Hepatic transcriptomic pathway analysis in ob/ob mice under NCD or FFD regimes. (A) Heat map visualization of biological categories related to NASH disease progression for 30 weeks. Values are expressed as–log (P-values). (B) Top 50 canonical pathways at 18 weeks. Red stars, green stars, purple stars, and blue stars indicate pathways involved in lipid metabolism, inflammatory processes, fibrosis and cancer and oxidative stress, respectively.

  • Fig. 6.
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    Fig. 6.

    Hepatic gene expression analysis in ob/ob mice under NCD or FFD regimes for 2, 4, 8, 12, 18, 24 and 30 weeks. (A) Heat map of genes involved in inflammation and fibrosis. Red, upregulated; green, downregulated. (B–F) Expression levels of selected genes. Values are mean±s.e.m., n=5. *P<0.05, **P<0.01 versus NCD (Student's t-test, two-tailed).

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Keywords

  • Fast food
  • Fibrosis
  • Non-alcoholic steatohepatitis
  • Hyperinsulinemia
  • Obesity

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Research Article
Longitudinal characterization of diet-induced genetic murine models of non-alcoholic steatohepatitis with metabolic, histological, and transcriptomic hallmarks of human patients
Naomichi Abe, Sayuka Kato, Takuma Tsuchida, Kanami Sugimoto, Ryuta Saito, Lars Verschuren, Robert Kleemann, Kozo Oka
Biology Open 2019 8: bio041251 doi: 10.1242/bio.041251 Published 1 May 2019
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Research Article
Longitudinal characterization of diet-induced genetic murine models of non-alcoholic steatohepatitis with metabolic, histological, and transcriptomic hallmarks of human patients
Naomichi Abe, Sayuka Kato, Takuma Tsuchida, Kanami Sugimoto, Ryuta Saito, Lars Verschuren, Robert Kleemann, Kozo Oka
Biology Open 2019 8: bio041251 doi: 10.1242/bio.041251 Published 1 May 2019

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