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

作者：Klein Sabine; Kloesel Jeremias; Schierwagen Robert; Koerner Christian; Granzow Michaela; Huss Sebastian; Mazar Irela Gretchen Reza; Weber Susanne; van den Ven Peter F M; Pieper Fuerst Ursula; Fuerst Dieter O; Nattermann Jacob; Lammert Frank; Sauerbruch Tilman; Trebicka Jonel^*

来源：Laboratory Investigation, 2012, 92(10): 1440-1450.

DOI：10.1038/labinvest.2012.106

• cytokines
• fibrogenesis
• hepatic stellate cells
• HMG-CoA reductase inhibitor
• myofibroblasts
• senescence

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-beta 1, connective tissue growth factor and TIMP1) and procollagen la were analyzed by real time PCR. Proliferation was investigated by 5%26apos;-bromo-2%26apos;-deoxyuridine assays. alpha-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 beta-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 beta-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. Laboratory Investigation (2012) 92, 1440-1450; doi:10.1038/labinvest.2012.106; published online 13 August 2012

• 出版日期2012-10