Background & Aims
Activation of hepatic stellate cells (HSC) and transdifferentiation to myofibroblasts following liver injury is the main culprit for hepatic fibrosis. Myofibroblasts show increased proliferation, migration, contraction, and production of extracellular matrix (ECM). In vitro, HMG-CoA reductase inhibitors (statins) inhibit proliferation and induce apoptosis of myofibroblastic HSC. To investigate the antifibrotic effects of atorvastatin in vivo we used bile duct ligated rats (BDL).
BDL rats were treated with atorvastatin (15 mg/kg/d) immediately after ligation (prophylactically) or in on-going fibrosis (therapeutically). Fibrosis was assessed by hydroxyproline content and Sirius-red staining. The activation of HSC was investigated by analysis of αSMA expression. mRNA levels of cytokines and procollagen were analyzed by RT-PCR, and MMP-2 activity by zymography. Proliferation was assessed by expression of cathepsins (B and D), proliferating cell nuclear antigen (PCNA), and Ki67-staining. Apoptosis was characterized by caspase-3 activity, cleavage of PARP-1, and TUNEL assay. Hepatic inflammation was investigated by serum parameters and liver histology.
Prophylactic and early therapy with atorvastatin significantly attenuated fibrosis and HSC activation. Later therapy lacked significant effects on fibrosis but reduced profibrotic cytokine expression and led to a more quiescent state of HSC with less proliferation and apoptosis, while hepatic inflammation did not change.
This study shows that very early atorvastatin treatment inhibits HSC activation and fibrosis in the BDL model in vivo, while late treatment reduces HSC turnover and activity. Our findings underline that long-term studies in humans are warranted.
Abbreviations:HSC (hepatic stellate cells), ECM (extracellular matrix), TGFβ (transforming growth factor-β), CTGF (connective tissue growth factor), PDGFβ-R (platelet-derived growth factor-β receptor), HMG-CoA-R (3-hydroxy-3-methylglutaryl-coenzyme-A-reductase), BDL (bile duct ligation), PCNA (proliferating cell nuclear antigen), PARP-1 (poly (ADP ribose) polymerase), ELISA (enzyme-linked immunosorbent assay), AST (aspartate aminotransferase), ALT (alanine aminotransferase), HCl (hydrogen chloride), αSMA (α-smooth muscle actin), SDS–PAGE (sodium dodecyl sulfate–polyacrylamide gel electrophoresis), RT-PCR (real-time polymerase chain reaction), CT-value (number of cycles), SEM (standard error of the mean), MMP-2 (matrix metalloproteinase-2), GAPDH (glyceraldehydes-3-P dehydrogenase)
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Published online: July 01, 2010
Accepted: April 15, 2010
Received in revised form: March 23, 2010
Received: November 6, 2009
© 2010 European Association for the Study of the Liver. Published by Elsevier Inc. All rights reserved.