Establishment of a murine model of acute-on-chronic liver failure with multi-organ dysfunction

Acute-on-chronic liver failure (ACLF) is a distinct clinical entity with high probability of organ failure and mortality. Since patients generally present late, experimental models are needed to understand the pathophysiology and natural course of the disease.To reproduce the syndrome of ACLF, chronic liver disease was induced in C57BL6 mice (6-8 weeks; approximately 20-24 g weight) by intraperitoneal administration of carbon tetrachloride (CCl4) for 10 weeks followed by an acute injury with acetaminophen (APAP) and lipopolysaccharide (LPS). Blood, ascitic fluid, and organs were collected to study cell death, regeneration, and fibrosis.At 24 h post-APAP/LPS infusion, the liver tissue showed increased hepatocyte ballooning and endothelial cell TUNEL positivity. This was followed by progressive hepatocyte necrosis from perivascular region at day 7 to lobular region by day 11. ACLF (day 7 and day 11) animals showed increase in bilirubin (p < 0.05), prothrombin time (p < 0.0001), blood ammonia (p < 0.001), and portal pressure post-acute hepatocellular injury similar to human ACLF. Ascites was noticed by day 11 with median serum-ascites albumin gradient of 1.2 (1.1-1.3) g/dL. In comparison to cirrhosis, ACLF group (day 7 and day 11) showed significant decrease in Sirius red (p ≤ 0.0001), collagen1 (p < 0.0001), and a-SMA proportionate area (p < 0.0001) with loss of hepatocytes regeneration (p < 0.005). At day 11, ACLF animals also showed significant increase in serum creatinine (p < 0.05) and acute tubular necrosis suggestive of organ failure, compared to cirrhotic animals.The CCL4/APAP/LPS (CALPS) model of ACLF mimics the clinical, biochemical, and histological features of ACLF with demonstrable progressive hepatocellular necrosis, liver failure, impaired regeneration, development of portal hypertension, and organ dysfunction in an animal with chronic liver disease.