Treatment started 1 week after development of ascites and stopping CCl4 administration

in a setting of advanced cirrhosis or after 2 weeks of BDL, in a precirrhotic stage. Experiments were performed 1 hour after the last dose of terutroban or vehicle. Treatments were prepared by a third person and experimental studies were realized blindly. The code was kept sealed until the final analysis of the results. find protocol The dose of terutroban used has been previously shown to have antivasoconstricting and antiatherosclerotic properties.[16, 22, 23] The animals were kept in environmentally controlled animal facilities at the Institut d’Investigacions Biomèdiques August Pi i Sunyer. All procedures were approved by the Laboratory Animal Care and Use Committee of the University of Barcelona and were conducted in accordance with European Community guidelines for the protection of animals used for experimental and other scientific purposes (EEC Directive 86/609). Cirrhotic rats were anesthetized with intraperitoneal ketamine hydrochloride (100 mg/kg; Merial Laboratories, Barcelona, Spain) plus midazolam (5 mg/kg intraperitoneally; Laboratorios Reig Jofré, Barcelona, Spain). The femoral artery and the ileocolic vein were cannulated with PE-50 catheters to measure mean arterial pressure

(MAP; mmHg) and portal pressure (PP; mmHg), respectively. Perivascular ultrasonic transit-time flow probes connected to a flow meter (Transonic Systems, Ithaca, NY) were placed around the portal vein, as close as possible to the liver to measure portal blood flow perfusing the Roscovitine nmr liver (PBF; mL/min/g liver) and around the superior mesenteric artery, in BDL cirrhotic rats, to measure superior mesenteric artery blood flow (SMABF, mL/min/100g body weight). Hepatic vascular resistance (HVR, mmHg/mL/min/g liver) was calculated as: PP/PBF; and superior mesenteric artery resistance (SMAR, mmHg/mL/min/100g

body weight) was calculated as (MAP-PP)/SMABF. Blood pressures and flows were registered on a multichannel computer-based recorder (PowerLab; AD Instruments, Colorado Springs, CO). The temperature of the animals was RNA Synthesis inhibitor maintained at 37 ± 0.5°C. Hemodynamic data were collected after a 20-minute stabilization period. To determine if terutroban correctly blocked the TP receptor in a subgroup of CCl4 and BDL cirrhotic rats (n = 3) treated with terutroban (30 mg/kg) or vehicle for 2 weeks, measurements of MAP and PP were performed before and after the intravenous infusion of 10 μg/kg U46619.[24] U46619 (9,11-dideoxy-9α,11α-methanoepoxy-prosta-5Z,13E-dien-1-oic acid; Cayman Chemical, Tallin, Estonia) is a synthetic TXA2 analog that specifically activates the TP-receptor. An additional group of cirrhotic rats were randomized to receive terutroban (30 mg/kg; n = 8 in CCl4; n = 8 in BDL) or vehicle (n = 9 in CCl4; n = 9 in BDL) for 3 days.