Nidhi Nautiyal1,2, Deepanshu Maheshwari1, Dinesh Mani Tripathi1, Dhananjay Kumar1, Rekha Kumari1, Suchi Gupta3, Sachin Sharma2, Sujata Mohanty3, Anupama Parasar1, Chhagan Bihari4, Subhrajit Biswas2, Archana Rastogi4, Rakhi Maiwall5, Anupam Kumar6, Shiv Kumar Sarin7,8. 1. Department of Molecular and Cellular Medicine, Institute of Liver and Biliary Sciences (ILBS), New Delhi, 110 070, India. 2. Amity Institute of Molecular Medicine and Stem Cell Research, Amity University, Greater Noida, India. 3. All India Institute of Medical Sciences, New Delhi, India. 4. Department of Pathology, Institute of Liver and Biliary Sciences, New Delhi, India. 5. Department of Hepatology, Institute of Liver and Biliary Sciences (ILBS), New Delhi, 110 070, India. 6. Department of Molecular and Cellular Medicine, Institute of Liver and Biliary Sciences (ILBS), New Delhi, 110 070, India. dr.anupamkumar.ilbs@gmail.com. 7. Department of Molecular and Cellular Medicine, Institute of Liver and Biliary Sciences (ILBS), New Delhi, 110 070, India. shivsarin@gmail.com. 8. Department of Hepatology, Institute of Liver and Biliary Sciences (ILBS), New Delhi, 110 070, India. shivsarin@gmail.com.
Abstract
BACKGROUND AND AIMS: 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. METHODOLOGY: 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. RESULTS: 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. CONCLUSION: 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.
BACKGROUND AND AIMS: 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. METHODOLOGY: 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. RESULTS: 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. CONCLUSION: 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.