Atorvastatin inhibits proliferation and apoptosis, but induces senescence in hepatic myofibroblasts and thereby attenuates hepatic fibrosis in rats

• Published in: Laboratory investigation Vol. 92; no. 10; pp. 1440 - 1450
• Main Authors: Klein, Sabine, Klösel, Jeremias, Schierwagen, Robert, Körner, Christian, Granzow, Michaela, Huss, Sebastian, Mazar, Irela Gretchen Reza, Weber, Susanne, van den Ven, Peter F M, Pieper-Fürst, Ursula, Fürst, Dieter O, Nattermann, Jacob, Lammert, Frank, Sauerbruch, Tilman, Trebicka, Jonel
• Format: Journal Article
• Language: English
• Published: New York Elsevier Inc 01.10.2012; Nature Publishing Group US; Nature Publishing Group
• Subjects: 631/80/509; 631/92/609; 692/699/1503/1607/1605; Actins - immunology; Actins - metabolism; Aging - drug effects; Animals; Apoptosis - drug effects; Atorvastatin; beta-Galactosidase - analysis; beta-Galactosidase - metabolism; Biological and medical sciences; Biotechnology; Cell Proliferation - drug effects; Collagen - metabolism; Connective Tissue Growth Factor - genetics; Connective Tissue Growth Factor - metabolism; cytokines; Desmin - immunology; Desmin - metabolism; Dose-Response Relationship, Drug; fibrogenesis; Flow Cytometry; Fundamental and applied biological sciences. Psychology; Gastroenterology. Liver. Pancreas. Abdomen; hepatic stellate cells; Hepatic Stellate Cells - drug effects; Heptanoic Acids - pharmacology; HMG-CoA reductase inhibitor; Hydroxymethylglutaryl-CoA Reductase Inhibitors - pharmacology; Investigative techniques, diagnostic techniques (general aspects); Laboratory Medicine; Liver - cytology; Liver - drug effects; Liver - metabolism; Liver Cirrhosis - drug therapy; Liver. Biliary tract. Portal circulation. Exocrine pancreas; Male; Medical sciences; Medicine; Medicine & Public Health; myofibroblasts; Myofibroblasts - drug effects; Other diseases. Semiology; Pathology; Pyrroles - pharmacology; Rats; Rats, Sprague-Dawley; research-article; senescence; Tissue Inhibitor of Metalloproteinase-1 - genetics; Tissue Inhibitor of Metalloproteinase-1 - metabolism; Transforming Growth Factor beta1 - genetics; Transforming Growth Factor beta1 - metabolism; senescence; hepatic stellate cells; cytokines; fibrogenesis; myofibroblasts; HMG-CoA reductase inhibitor; Cell proliferation; Biotechnology; Senescence; Rat; Liver; Myofibroblast; Hepatic disease; Hypocholesterolemic agent; Clinical biology; Hepatic fibrosis; Digestive system; Enzyme; Rodentia; Cytokine; Enzyme inhibitor; Statin derivative; Kupffer cell; Vertebrata; Mammalia; Cell death; Animal; Atorvastatin; Digestive diseases; Hydroxymethylglutaryl-CoA reductase; Oxidoreductases; Apoptosis; Antilipemic agent
• ISSN: 0023-6837; 1530-0307
• DOI: 10.1038/labinvest.2012.106

Hepatic myofibroblasts (MFB) show increased proliferation, migration and collagen production, which are crucial for hepatic fibrogenesis. Atorvastatin treatment inhibits proliferation, apoptosis and cytokine production of MFB in bile duct-ligated (BDL) rats in vivo. Here, we have further investigated the underlying mechanisms. Primary rat hepatic stellate cells (HSC) were isolated and culture-activated to hepatic MFB. Following 3 days of incubation with atorvastatin (10−4, 10−5 and 10−6 M), transcription levels of profibrotic cytokines (transforming growth factor-β1, connective tissue growth factor and TIMP1) and procollagen Ia were analyzed by real time PCR. Proliferation was investigated by 5'-bromo-2'-deoxyuridine assays. α-Smooth muscle actin protein expression was examined by western blotting. Fluorescence-activated cell sorting analysis of Annexin V and propidium iodide were used to measure apoptosis. Furthermore, p21 western blotting and β-galactosidase staining were investigated in MFB as senescence markers. Subsequently, hepatic expression of desmin and senescence markers were analyzed in the livers of rats receiving atorvastatin (15 mg/kg*d) for 1 week starting 3 and 5 weeks after BDL. Atorvastatin inhibited the activation of HSC to MFB and decreased cytokine and collagen production in MFB in vitro. In addition, proliferation, cytokine and collagen production of MFB were reduced by atorvastatin. Atorvastatin initiated apoptosis at 10−4 M and attenuated it at 10−5 M. Atorvastatin induced p21 protein expression and β-galactosidase staining of MFB in vitro and in vivo. Atorvastatin elicits similiar effects on MFB as previously seen in vivo: it decreases MFB turnover and fibrogenesis. We suggest that a further mechanism explaining these effects is senescence of cells.