The presence of ascites was confirmed at the time the rats were killed following laparotomy by abdominal fluid drainage. Diagnosis of cirrhosis was confirmed postmortem by microscopic examination of hematoxylin-stained liver sections. A
43% albumin (Sigma Aldrich, Milan, Italy) solution in 0.9% saline was injected into the caudal vein of 15 rats with cirrhosis and ascites and 15 control rats. Two albumin doses were administered: the first dose (1.5 g/kg) was infused 3 days before sacrifice of the animals, and the second dose (1 g/kg) was infused the day before sacrifice. The doses were chosen in order to simulate those which are commonly given to patients with cirrhosis and ascites when a spontaneous bacterial peritonitis (SBP) is diagnosed in order to prevent the development
Ensartinib concentration of renal failure.10, Selleck Panobinostat 11 Fifteen control animals and 15 rats with cirrhosis and ascites were treated with an equivalent volume of 0.9% of saline. A synthetic plasma expander, hydroxyethyl starch (HES) (10% hydroxyethyl starch HES 200/0.5 in isotonic sodium chloride solution, Fresenius Kabi Deutschland, Friedberg, Germany) was injected into the caudal vein of 15 rats with cirrhosis and ascites and 15 control rats. HES was administered at the same doses and with the same schedule of albumin. To monitor the effects of albumin, saline, or HES on the effective circulating volume a blood sample was obtained in rats with cirrhosis and ascites just before and after administration of the two doses of each plasma expander for the measurement of plasma renin activity (PRA). PRA was measured by means of RIA (Radim, Pomezia, Italy). After intraperitoneal injection of 300 μL of heparin (5000 U/mL) animals were sacrificed by decapitation. Subsequently, the heart, cannulated through the aorta, was mounted in a Langendorff apparatus for retrograde perfusion, perfused at constant flow (10 mL/min), and electrically driven at a frequency of
6 Hz using platinum electrodes placed in the left atrium as described.17 Left ventricular developed pressure (LVDP) was measured by inserting a steel cannula into the left ventricle and connecting it to a pressure transducer (2B Instruments, Besozzo, Italy). The perfusion medium was a modified Krebs-Henselheit MCE公司 saline solution with the following composition: 118 mM NaCl, 4.7 mM KCl, 2.5 mM CaCl2, 1.2 mM MgSO4, 25 mM NaHCO3, 11.1 mM glucose, and 2.0 mM disodium pyruvate, bubbled with a 95%/5% O2/CO2 mixture at 37°C. Following a stabilizing period of 20 minutes, the heart was stimulated with increasing concentrations of isoproterenol (from 10−10 to 10−8 M) to obtain a concentration-response curve. The LVDP was continuously recorded and stored by a real-time digital acquisition and analysis system (model MP-100, Biopac System, Santa Barbara, CA). LVDP was calculated as the difference between systolic and diastolic values of LV pressure.