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

Nidhi Nautiyal,Rakhi Maiwall,Sujata Mohanty,Archana Rastogi,Dinesh Mani Tripathi,Rekha Kumari,Sachin Sharma,Suchi Gupta,Anupama Parasar,Subhrajit Biswas,Deepanshu Maheshwari,Chhagan Bihari,Dhananjay Kumar,Shiv Kumar Sarin,Anupam Kumar

doi:10.1007/s12072-021-10244-0

Abstract

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 (6-8weeks; approximately 20-24g weight) by intraperitoneal administration of carbon tetrachloride (CCl4) for 10weeks 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 24h 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.

Highlights

Acute-on-chronic liver failure (ACLF) is an increasingly recognized clinical entity encompassing an acute liver failure in patients with underlying chronic liver disease or cirrhosis, resulting in cascade of event leading to sepsis, multi-organ failure (MOF) and death [1,2,3,4,5,6]
Methodology and Results: To mimic the syndrome of ACLF, chronic liver disease was induced by intraperitoneal administration of carbon tetrachloride (CCl4) for 10 weeks, followed by acute injury with acetaminophen (APAP) and lipopolysaccharide (LPS) administration
We developed a mouse model of ACLF by chronic administration of CCl4 to produce chronic liver injury/cirrhosis followed by an acute insult with acetaminophen (APAP) and a low dose of lipopolysaccharide (LPS); the CALPS model, to mimic the situation of an acute insult on a chronic liver disease along with introduction of intestinal endotoxemia

Summary

Introduction

Acute-on-chronic liver failure (ACLF) is an increasingly recognized clinical entity encompassing an acute liver failure in patients with underlying chronic liver disease or cirrhosis, resulting in cascade of event leading to sepsis, multi-organ failure (MOF) and death [1,2,3,4,5,6]. One of the major challenges in the ACLF is the poor understanding of underlying pathophysiology leading to rapid progression of liver injury, sepsis and MOF. A representative experimental model that mimics the course of human ACLF is crucial to decipher the pathophysiological journey of the disease and development of new therapeutic strategies for improved clinical outcomes for ACLF patients. Experimental models of ACLF are needed to understand the pathophysiology and natural course of the disease

Methods

Results

Conclusion