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

BMC Gastroenterology

Yi-Tsau Huang³  Jaw-Ching Wu⁴  Kuo-Ching Yang⁷  Yung-Tsung Chiu²  Yun Lin⁶  Ting-Fang Lee¹  Yi-Chao Hsu⁵  Lee-Won Chong⁷ 

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

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

Others  :  1137276
DOI  :  10.1186/s12876-015-0248-8

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

【 摘 要 】

Background

Non-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).

Methods

Palmitate (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 gp91^phox 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.

Results

In vitro, Flu (1–20 μM) inhibited PA-induced free-radical production, gp91^phox 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.

Conclusions

We 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】

   
2015 Chong et al.; licensee BioMed Central.

【 图 表 】

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