We hypothesized that PBEF might play a role selleck chemical in acute and chronic liver damage. We show that PBEF is strongly up-regulated in human chronic liver diseases and acute experimental hepatitis. Mice overexpressing hepatic
PBEF at baseline are more susceptible to liver damage during ConA- or D-galactosamine/lipopolysaccharide (LPS)–mediated hepatitis, whereas FK866 protected mice from acute hepatic injury induced by either ConA or D-galactosamine/LPS. ALT, alanine aminotransferase; AST, aspartate aminotransferase; ConA, concanavalin A; CTP, Child-Turcotte-Pugh; ELISA, enzyme-linked immunosorbent assay; IFNγ, interferon-γ; IL, interleukin; LPS, lipopolysaccharide; LTA, lipoteichoic acid; mRNA, messenger RNA; NAD, nicotinamide adenine dinucleotide; Nampt, nicotinamide phosphoribosyltransferase; PARP, poly (adenosine diphosphate-ribose) polymerase; PBEF, pre–B cell colony–enhancing factor; PCR, polymerase chain reaction; RT-PCR, reverse-transcription polymerase chain reaction; shRNA, short hairpin RNA; SIRT, sirtuin; TNFα, tumor necrosis factor α; TUNEL, terminal deoxynucleotidyl transferase–mediated deoxyuridine triphosphate
nick-end labeling. Serum samples were obtained from 83 randomly selected, consecutive patients with clinically, biochemically, radiologically, and histologically confirmed diagnosis of chronic liver disease. Chronic liver disease was staged according to Child-Turcotte-Pugh (CTP) criteria.20 Thirty-nine age- and sex-matched healthy subjects served as a control group. Baseline characteristics of chronic liver disease patients are reported in Table 1. Informed consent was obtained, and the study was approved by the local CP-690550 solubility dmso ethics committee of the Innsbruck Medical University. Nine milliliters of blood were collected in Sarstedt Monovette tubes. Blood was centrifuged at 1,200g for 15 minutes and 1-mL aliquots were stored at −80°C until assayed. Each sample was assigned an encoding number, and all assays were performed in duplicate in a blinded manner. Six- to eight-week old MCE公司 female C57BL/6 mice were obtained from Charles River (Sulzfeld, Germany). Mice were housed in accordance with institutional animal care with open access
to standard chow and water. Animal experiments were approved by the Austrian Federal Ministry of Science and Research (license number: 66011/34-II/10b/2009). Unless stated otherwise, mice were injected intravenously through the lateral tail vein with 15 mg/kg ConA from Canavalia ensiformis (Sigma-Aldrich, St. Louis, MO) in endotoxin-free phosphate-buffered saline. Mice received an intraperitoneal injection of 700 mg/kg D-galactosamine (Carl Roth, Karlsruhe, Germany) and 1 μg/kg lipopolysaccharide (InvivoGen, San Diego, CA). Mice were euthanized at the indicated time after injection. FK866 was purchased from Axon Medchem (Groningen, Netherlands) and dissolved in dimethyl sulfoxide; 25-mg/mL aliquots were stored at −80°C until further use.