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

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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.

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Abbreviations

ACLF:

Acute-on-chronic liver failure

CCl4 :

Carbon tetrachloride

APAP:

Acetaminophen

LPS:

Lipopolysaccharide

ATN:

Acute tubular necrosis

SAAG:

Serum-ascites albumin gradient

SIRS:

Systemic inflammatory response

TUNEL:

Terminal deoxynucleotidyl transferase dUTP nick end labeling

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Acknowledgements

This work was supported by Science and Engineering Research Board (SERB), Government of India Grant IR/SB/EF/02/2016.

Funding

This work was supported by Science and Engineering Research Board (SERB), Government of India Grant IR/SB/EF/02/2016.

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Authors and Affiliations

  1. Department of Molecular and Cellular Medicine, Institute of Liver and Biliary Sciences (ILBS), New Delhi, 110 070, India

    Nidhi Nautiyal, Deepanshu Maheshwari, Dinesh Mani Tripathi, Dhananjay Kumar, Rekha Kumari, Anupama Parasar, Anupam Kumar & Shiv Kumar Sarin

  2. Department of Hepatology, Institute of Liver and Biliary Sciences (ILBS), New Delhi, 110 070, India

    Rakhi Maiwall & Shiv Kumar Sarin

  3. Department of Pathology, Institute of Liver and Biliary Sciences, New Delhi, India

    Chhagan Bihari & Archana Rastogi

  4. Amity Institute of Molecular Medicine and Stem Cell Research, Amity University, Greater Noida, India

    Nidhi Nautiyal, Sachin Sharma & Subhrajit Biswas

  5. All India Institute of Medical Sciences, New Delhi, India

    Suchi Gupta & Sujata Mohanty

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  1. Nidhi Nautiyal
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  2. Deepanshu Maheshwari
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Contributions

NN, AK, and SKS contributed to the design of the studies. NN performed the majority of experiments, with assistance from MD, KD, KR, and PA. DMT and NN contributed in hemodynamics assessment. GS and SM help in RT-PCR. SS helped in western blot. CB and RA help in histopathology analysis. AK, RM, SB, and SKS assisted with interpretation of the findings. The manuscript was written by NN and AK with critical input from SKS and RM.

Corresponding authors

Correspondence to Anupam Kumar or Shiv Kumar Sarin.

Ethics declarations

Ethics approval

Animal care and all experimental procedures were approved by the Institutional Animal Ethics Committee (IAEC) under the project approval (IAEC/ILB/17/03) from Department of Science and Technology, Government of India.

Conflict of interest

Nautiyal Nidhi, Maheshwari Deepanshu, Tripathi Dinesh Mani, Kumar Dhananjay, Kumari Rekha, Gupta Suchi, Sharma Sachin, Mohanty Sujata, Parasar Anupama, Bihari Chhagan, Biswas Subhrajit, Rastogi Archana, Maiwall Rakhi, Kumar Anupam, Shiv Kumar Sarin have no conflict of interest.

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Nautiyal, N., Maheshwari, D., Tripathi, D.M. et al. Establishment of a murine model of acute-on-chronic liver failure with multi-organ dysfunction. Hepatol Int 15, 1389–1401 (2021). https://doi.org/10.1007/s12072-021-10244-0

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