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The intrahepatic expression levels of bile acid transporters are inversely correlated with the histological progression of nonalcoholic fatty liver disease

  • Original Article—Liver, Pancreas, and Biliary Tract
  • Published:
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Abstract

Background

Nonalcoholic fatty liver disease (NAFLD) presents as a spectrum ranging from simple steatosis to nonalcoholic steatohepatitis (NASH). The latter is progressive, and its pathogenesis remains poorly understood. Recently, bile acid (BA) metabolism has become a therapeutic focus in NASH patients. The aim of the present study was to explore changes in bile acid metabolism in NAFLD patients in the context of disease progression.

Methods

We prospectively enrolled patients with clinically suspected NAFLD. Patients taking ursodeoxycholic acid were excluded. The intrahepatic expression levels of genes associated with BA metabolism were determined by quantitative PCR and immunohistochemistry.

Results

Seventy-eight patients (male:female = 49:29) histologically diagnosed with NAFLD were analyzed. The expression levels of farnesoid X receptor, liver receptor homolog 1, and small heterodimer partner, key proteins in BA synthesis, significantly decreased as the NAFLD activity score (NAS) increased in either males or females. The levels of cholesterol 7 alpha-hydroxylase, the rate-limiting enzyme of BA synthesis, were not changed. Notably, the expression levels of a main export transporter, bile salt export pump (BSEP), significantly decreased as the NAS and the each NAS component increased in both genders. The decreases of BSEP levels were also observed by immunohistochemistry, particularly in areas with pronounced fatty changes in cases with high NAS.

Conclusions

The expression levels of the BA export transporter BSEP were inversely correlated with NAS in NAFLD patients. Such down-regulation may cause excessive BA levels in hepatocytes, leading to cell injury. Our findings may afford new insights into the pathogenesis of NASH.

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Abbreviations

NAFLD:

Nonalcoholic fatty liver disease

NASH:

Nonalcoholic steatohepatitis

BA:

Bile acid

FXR:

Farnesoid X receptor

AST:

Aspartate aminotransferase

ALT:

Alanine aminotransferase

UDCA:

Ursodeoxycholic acid

OGTT:

Oral glucose tolerance test

CRP:

C-reactive protein

GGT:

Gamma-glutamyl transpeptidase

PT-INR:

Prothrombin time-international normalized ratio

FBG:

Fasting blood glucose

HDL:

High-density lipoprotein

LDL:

Low-density lipoprotein

TG:

Triglyceride

NAS:

NAFLD activity score

BSEP:

Bile salt export pump

MRP2:

Multidrug resistance-associated protein 2

mRNA:

Messenger RNA

SHP:

Small heterodimer partner

LRH1:

Liver receptor homolog 1

NTCP:

Na+/taurocholate cotransporter

CYP7A1:

Cholesterol 7 alpha-hydroxylase

GAPDH:

Glyceraldehyde-3-phosphate dehydrogenase

PFIC2:

Progressive familial intrahepatic cholestasis type 2

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Acknowledgments

We thank Ms. Seiko Shinzawa (Department of Gastroenterology, Graduate School of Medicine, the University of Tokyo) for kind advice on research procedures. We thank Mr. Takeshi Shimamoto (Kameda Medical Center Makuhari) for detailed assistance with statistics. We also thank Mr. Kei Sakuma (Department of Pathology, Graduate School of Medicine, the University of Tokyo) for processing specimens for immunohistochemical analysis. This work was supported by Health Sciences Research Grants from the Ministry of Health, Labor, and Welfare of Japan (Research on Hepatitis) and AMED (the Japan Agency for Medical Research and Development). No additional external funding was received. The funders played no role in study design, data collection, or analysis, the decision to publish, or manuscript preparation.

Authors’ contributions

KO, TT, and KE contributed to the study concept and design, acquisition of data, analysis and interpretation of data, and drafting of the manuscript. HF, AK, KM, and HY participated in critical revision of the manuscript to ensure that the intellectual content was of a high standard. JS and MF contributed to histological analysis. KK participated in study concept and design, analysis and interpretation of data, drafting of the manuscript, critical revision of the manuscript to ensure that the intellectual content was of a high standard, and study supervision. We confirm that all authors have reviewed and approved of the final version of the manuscript.

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Authors and Affiliations

  1. Department of Gastroenterology, Graduate School of Medicine, The University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo, 113-8655, Japan

    Kazuya Okushin, Kenichiro Enooku, Hidetaka Fujinaga, Akira Kado & Kazuhiko Koike

  2. Department of Infectious Diseases, Graduate School of Medicine, The University of Tokyo, Tokyo, Japan

    Takeya Tsutsumi & Hiroshi Yotsuyanagi

  3. Department of Pathology, Graduate School of Medicine, The University of Tokyo, Tokyo, Japan

    Junji Shibahara & Masashi Fukayama

  4. Department of Infection Control and Prevention, Graduate School of Medicine, The University of Tokyo, Tokyo, Japan

    Kyoji Moriya

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  1. Kazuya Okushin
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Correspondence to Kazuhiko Koike.

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Supplementary Fig. 1

The associations between the changes of the expression levels of BA transporters and each component of the NAS in females. A, B, and C: Graded by steatosis. Significant associations were not evident. D, E, and F: Graded by lobular inflammation. BSEP and NTCP were significantly downregulated with progressions of lobular inflammation. G, H, and I: Graded by hepatocyte ballooning. Significant associations were not evident. (TIFF 71 kb)

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Okushin, K., Tsutsumi, T., Enooku, K. et al. The intrahepatic expression levels of bile acid transporters are inversely correlated with the histological progression of nonalcoholic fatty liver disease. J Gastroenterol 51, 808–818 (2016). https://doi.org/10.1007/s00535-015-1148-y

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  • DOI: https://doi.org/10.1007/s00535-015-1148-y

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