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A high-cholesterol diet promotes steatohepatitis and liver tumorigenesis in HCV core gene transgenic mice

  • Genotoxicity and Carcinogenicity
  • Published:
Archives of Toxicology Aims and scope Submit manuscript

A Correction to this article was published on 01 June 2019

This article has been updated

Abstract

Previous epidemiological studies have suggested a link between high-cholesterol intake and liver disease progression, including hepatocellular carcinoma (HCC). However, the precise mechanism of hepatotoxicity and hepatocarcinogenesis caused by excessive cholesterol consumption remains unclear. We aimed to investigate the impact of dietary cholesterol using hepatitis C virus core gene transgenic (HCVcpTg) mice, which spontaneously developed HCC with age. Male HCVcpTg mice were treated for 15 months with either a control diet or an isocaloric diet containing 1.5% cholesterol, and liver phenotypes and tumor-associated signaling pathways were evaluated. The high-cholesterol diet-fed HCVcpTg mice exhibited a significantly higher incidence of liver tumors compared with the control diet mice (100% vs. 41%, P < 0.001). The diet induced steatohepatitis with pericellular fibrosis and evoked higher mRNA expression of pro-inflammatory and pro-fibrotic mediators along with enhanced hepatocyte proliferation and greater oxidative and endoplasmic reticulum stress in the liver. Moreover, long-term consumption of cholesterol-rich diet activated nuclear factor-kappa B (NF-κB) and p62/sequestosome 1 (Sqstm1)-nuclear factor erythroid 2 (NRF2) axis, enhanced fibrogenesis, and consequently accelerated hepatic tumorigenesis. In conclusion, these results demonstrate that a high-cholesterol diet facilitates liver tumorigenesis by inducing steatohepatitis, promoting hepatocyte division, and up-regulating cellular stress and pro-inflammatory NF-κB and detoxifying p62/Sqstm1-NRF2 signals. Therefore, high dietary cholesterol should be avoided in HCV-infected patients to prevent development of steatohepatitis, liver fibrosis, and HCC.

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  • 08 June 2019

    In the original publication of the article.

Abbreviations

4-HNE:

4-Hydroxy-nonenal

ALT:

Alanine aminotransferase

AST:

Aspartate aminotransferase

ALP:

Alkaline phosphatase

αSMA:

α-Smooth muscle actin

BAX:

B-cell leukemia/lymphoma 2-associated X protein

CHOP:

CCAAT/enhancer-binding protein homologous protein

DR5:

Tumor necrosis factor receptor superfamily member 10b

ER:

Endoplasmic reticulum

FA:

Fatty acid

HCC:

Hepatocellular carcinoma

F-Chol:

Free cholesterol

GAPDH:

Glyceraldehyde-3-phosphate dehydrogenase

HCV:

Hepatitis C virus

HCVcpTg:

HCV core gene transgenic

HMG-CoA:

Hydroxymethylglutaryl-CoA

IL:

Interleukin

LXR:

Liver X receptor

NASH:

Non-alcoholic steatohepatitis

NEFA:

Non-esterified fatty acid

NF-κB:

Nuclear factor-kappa B

NQO1:

NAD(P)H quinone dehydrogenase 1

NRF2:

Nuclear factor erythroid 2

PC:

phospholipid

PCNA:

Proliferating cell nuclear antigen

PPAR:

Peroxisome proliferator-activated receptor

qPCR:

Quantitative polymerase chain reaction

SDS-PAGE:

Sodium dodecyl sulfate-polyacrylamide gel electrophoresis

SEM:

Standard error of the mean

Sqstm1:

Sequestosome 1

SREBP:

Sterol regulatory element-binding protein

T-BA:

Total bile acid

T-Chol:

Total cholesterol

TG:

Triglyceride

TGF-β1:

Transforming growth factor-β1

TLR:

Toll-like receptor

TNFα:

Tumor necrosis factor-α

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Acknowledgements

We appreciate Mr. Trevor Ralph for his English editorial assistance and Dr. Takero Nakajima and Dr. Ruan Guo (Shinshu University School of Medicine) for their invaluable help, advice, instruction, and encouragement. We thank Research Center for Supports to Advanced Science, Shinshu University, for technical supports and animal care. This study was partially supported by JSPS Grants-in-Aid for Scientific Research (KAKENHI Grant number 16K08734).

Funding

The authors have declared that no financial support exists.

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Authors and Affiliations

  1. Department of Metabolic Regulation, Shinshu University School of Medicine, Matsumoto, 390-8621, Japan

    Xiaojing Wang, Naoki Tanaka, Xiao Hu, Yu Lu, Fangping Jia & Toshifumi Aoyama

  2. Department of Gastroenterology, Lishui Hospital, Zhejiang University School of Medicine, Lishui, Zhejiang, People’s Republic of China

    Xiaojing Wang

  3. Research Center for Social Systems, Shinshu University, Matsumoto, Japan

    Naoki Tanaka

  4. Department of Pathophysiology, Hebei Medical University, Shijiazhuang, People’s Republic of China

    Xiao Hu

  5. Department of Gastroenterology, Shinshu University School of Medicine, Matsumoto, Japan

    Takefumi Kimura

  6. Department of Molecular Pathology, Shinshu University School of Medicine, Matsumoto, Japan

    Yoshiko Sato & Jun Nakayama

  7. Department of Infection Control and Prevention, The University of Tokyo, Tokyo, Japan

    Kyoji Moriya

  8. Department of Gastroenterology, The University of Tokyo, Tokyo, Japan

    Kazuhiko Koike

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  1. Xiaojing Wang
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  2. Naoki Tanaka
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Contributions

Design and writing the paper: NT and TA. Performing experiments: XW, NT, XH, TK, YL, FJ, and YS. Analyzing data: XW, NT, XH, TK, YL, and YS. Supervision: JN, KM, and KK. All authors read and approved the final manuscript.

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Correspondence to Naoki Tanaka.

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The original version of this article was revised: In the original publication of the article, there was a mistake in Fig. 2e and the author would like to correct it. It has now been corrected and replaced in the HTML and PDF versions.

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Wang, X., Tanaka, N., Hu, X. et al. A high-cholesterol diet promotes steatohepatitis and liver tumorigenesis in HCV core gene transgenic mice. Arch Toxicol 93, 1713–1725 (2019). https://doi.org/10.1007/s00204-019-02440-7

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