Fluvastatin attenuates hepatic steatosis-induced fibrogenesis in rats through inhibiting paracrine effect of hepatocyte on hepatic stellate cells

BMC Gastroenterology

Research Article

Yung-Tsung Chiu¹  Kuo-Ching Yang²  Yi-Chao Hsu³  Ting-Fang Lee⁴  Lee-Won Chong⁵  Jaw-Ching Wu⁶  Yun Lin⁷  Yi-Tsau Huang⁸ 

[1] Department of Medical Research and Education, Taichung Veterans General Hospital, Taichung, Taiwan;Division of Hepatology and Gastroenterology, Department of Internal Medicine, Shin Kong Wu Ho-Su Memorial Hospital, Taipei, Taiwan;Institute of Biomedical Sciences, Mackay Medical College, New Taipei City, Taiwan;Institute of Clinical Medicine, School of Medicine, National Yang Ming University, Taipei, Taiwan;Institute of Clinical Medicine, School of Medicine, National Yang Ming University, Taipei, Taiwan;Division of Hepatology and Gastroenterology, Department of Internal Medicine, Shin Kong Wu Ho-Su Memorial Hospital, Taipei, Taiwan;Institute of Clinical Medicine, School of Medicine, National Yang Ming University, Taipei, Taiwan;Translational Research Division, Medical Research Department, Taipei Veterans General Hospital, Taipei, Taiwan;National Research Institute of Chinese Medicine, No. 155-1, Li-Nong Street, Sec. 2, 112, Taipei, Taiwan;National Research Institute of Chinese Medicine, No. 155-1, Li-Nong Street, Sec. 2, 112, Taipei, Taiwan;Institute of Traditional Medicine, School of Medicine, National Yang Ming University, Taipei, Taiwan;

关键词: Non-alcoholic fatty liver disease;    Steatohepatitis;    Statins;    Fibrogenesis;    Hepatocyte;    Paracrine effect;   

DOI  :  10.1186/s12876-015-0248-8

 received in 2014-07-19, accepted in 2015-02-03,  发布年份 2015

来源: Springer

【 摘 要 】

BackgroundNon-alcoholic steatohepatitis (NASH) is associated with hepatic fibrogenesis. Despite well-known cholesterol-lowering action of statins, their mechanisms against NASH-mediated fibrogenesis remain unclear. This study aimed at investigating the in vitro and in vivo anti-fibrotic properties of fluvastatin (Flu).MethodsPalmitate (PA)-induced changes in intracellular hydrogen peroxide levels in primary rat hepatocytes (PRHs) and human hepatoma cell line (HepG2) were quantified by dichlorofluorescein diacetate (DCF-DA) dye assay, whereas changes in expressions of NADPH oxidase gp91phox subunit, α-smooth muscle actin (α-SMA), and NFκB p65 nuclear translocation were quantified with Western blotting. Quantitative real-time polymerase chain reaction (q-PCR) was used to investigate mRNA expressions of pro-inflammatory genes (ICAM-1, IL-6, TNF-α). Conditioned medium (CM) from PA-treated PRHs was applied to cultured rat hepatic stellate cell line, HSC-T6, with or without Flu-pretreatment for 2 h. Pro-fibrogenic gene expressions (COL1, TIMP-1, TGF-β1, α-SMA) and protein expression of α-SMA were analyzed. In vivo study using choline-deficient L-amino acid defined (CDAA) diet-induced rat NASH model was performed by randomly assigning Wistar rats (n = 28) to normal controls (n = 4), CDAA diet with vehicles, and CDAA diet with Flu (5 mg/kg or 10 mg/kg) (n = 8 each) through gavage for 4 or 8 weeks. Livers were harvested for histological, Western blot (α-SMA), and q-PCR analyses for expressions of pro-inflammatory (IL-6, iNOS, ICAM-1) and pro-fibrogenic (Col1, α-SMA, TIMP-1) genes.ResultsIn vitro, Flu (1–20 μM) inhibited PA-induced free-radical production, gp91phox expression, and NFκB p65 translocation in HepG2 and PRHs, while CM-induced α-SMA protein expression and pro-fibrogenic gene expressions in HSC-T6 were suppressed in Flu-pretreated cells compared to those without pretreatment. Moreover, α-SMA protein expression was significantly decreased in HSC-T6 cultured with CM from PA-Flu-treated PRHs compared to those cultured with CM from PA-treated PRHs. Flu also reduced steatosis and fibrosis scores, α-SMA protein expression, mRNA expression of pro-inflammatory and pro-fibrogenic genes in livers of CDAA rats.ConclusionsWe demonstrated PA-induced HSC activation through paracrine effect of hepatocyte in vitro that was significantly suppressed by pre-treating HSC with Flu. In vivo, Flu alleviated steatosis-induced HSC activation and hepatic fibrogenesis through mitigating inflammation and oxidative stress, suggesting possible therapeutic role of Flu against NASH.

【 授权许可】

Unknown   
© Chong et al.; licensee BioMed Central. 2015. This article is published under license to BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly credited. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.