Basic Study
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World J Hepatol. Sep 27, 2020; 12(9): 596-618
Published online Sep 27, 2020. doi: 10.4254/wjh.v12.i9.596
N-acetylcysteine and glycyrrhizin combination: Benefit outcome in a murine model of acetaminophen-induced liver failure
Charlotte Minsart, Sandrine Rorive, Arnaud Lemmers, Eric Quertinmont, Thierry Gustot
Charlotte Minsart, Arnaud Lemmers, Eric Quertinmont, Thierry Gustot, Laboratory of Experimental Gastroenterology, Erasme Hospital, Université Libre de Bruxelles, Brussels 1070, Belgium
Sandrine Rorive, Department of Pathology, Erasme Hospital, Université Libre de Bruxelles, Brussels 1070, Belgium
Sandrine Rorive, DIAPATH-Center for Microscopy and Molecular Imaging, Université Libre de Bruxelles, Gosselies 6041, Belgium
Arnaud Lemmers, Thierry Gustot, Department of Gastroenterology, Hepato Pancreatology and Digestive Oncology, Erasme Hospital, Université Libre de Bruxelles, Brussels 1070, Belgium
Thierry Gustot, Inserm Unité 1149, Centre de Recherche sur l’inflammation, Paris 75006, France
Thierry Gustot, UMR S_1149, Université Paris Diderot, Paris 75006, France
Author contributions: All authors contributed to the study conception and design; Material preparation, data collection and analysis were performed by Minsart C; Immunohistochemistry and histological analyses were carried out with the help of Rorive S; The first draft of the manuscript was written by Minsart C and Gustot T; all authors commented on previous versions of the manuscript, and all read and approved the final manuscript.
Supported by the Bourse du Conseil Médicalde l’hôpital Erasme, Fonds E. et S. Jacobs and Novartis Grant;The CMMI is supported by the European Regional Development Fund and Wallonia.
Institutional review board statement: This study did not involve humans and thus was exempt.
Institutional animal care and use committee statement: All animal experiments conformed to the internationally accepted principles for the care and use of laboratory animals. Animal protocols were approved by the local Ethic Committee of the Université Libre de Bruxelles (License No. 1230406, Animalerie de la Faculté de Médecine du Campus Erasme, Brussels, Belgium; Protocol Nos. 488N and 734N).
Conflict-of-interest statement: The authors declare they have no conflicts of interest in relation to this paper.
Data sharing statement: No additional data are available.
ARRIVE guidelines statement: The authors have read the ARRIVE guidelines, and the manuscript was prepared and revised according to the ARRIVE guidelines.
Open-Access: This article is an open-access article that was selected by an in-house editor and fully peer-reviewed by external reviewers. It is distributed in accordance with the Creative Commons Attribution NonCommercial (CC BY-NC 4.0) license, which permits others to distribute, remix, adapt, build upon this work non-commercially, and license their derivative works on different terms, provided the original work is properly cited and the use is non-commercial. See: http://creativecommons.org/licenses/by-nc/4.0/
Corresponding author: Charlotte Minsart, PhD, Research Scientist, Laboratory of Experimental Gastroenterology, Erasme Hospital, Université Libre de Bruxelles, 808, Route de Lennik, Brussels 1070, Belgium. charlotte.minsart@erasme.ulb.ac.be
Received: November 28, 2019
Peer-review started: November 28, 2019
First decision: December 30, 2019
Revised: June 29, 2020
Accepted: August 25, 2020
Article in press: August 25, 2020
Published online: September 27, 2020
Processing time: 297 Days and 20 Hours
Abstract
BACKGROUND

Acetaminophen overdose is the most frequent cause of drug-induced liver failure in developed countries. Substantial progress has been made in understanding the mechanism of hepatocellular injury, but N-acetylcysteine remains the only effective treatment despite its short therapeutic window. Thus, other hepatoprotective drugs are needed for the delayed treatment of acetaminophen-induced hepatotoxicity. Our interest focused on glycyrrhizin for its role as an inhibitor of high mobility group box 1 (HMGB1) protein, a member of the family of damage-associated molecular pattern, known to play an important pathological role in various diseases.

AIM

To investigate the efficacy of the N-acetylcysteine/glycyrrhizin combination compared to N-acetylcysteine alone in the prevention of liver toxicity.

METHODS

Eight-week-old C57BL/6J wild-type female mice were used for all our experiments. Mice fasted for 15 h were treated with acetaminophen (500 mg/kg) or vehicle (phosphate-buffered saline) by intraperitoneal injection and separated into the following groups: Glycyrrhizin (200 mg/kg); N-acetylcysteine (150 mg/kg); and N-acetylcysteine/glycyrrhizin. In all groups, mice were sacrificed 12 h following acetaminophen administration. The assessment of hepatotoxicity was performed by measuring plasma levels of alanine aminotransferase, aspartate aminotransferase and lactate dehydrogenase. Hepatotoxicity was also evaluated by histological examination of hematoxylin and eosin-stained tissues sections. Survival rates were compared between various groups using Kaplan-Meier curves.

RESULTS

Consistent with data published in the literature, we confirmed that intraperitoneal administration of acetaminophen (500 mg/kg) in mice induced severe liver injury as evidenced by increases in alanine aminotransferase, aspartate aminotransferase and lactate dehydrogenase but also by liver necrosis score. Glycyrrhizin administration was shown to reduce the release of HMGB1 and significantly decreased the severity of liver injury. Thus, the co-administration of glycyrrhizin and N-acetylcysteine was investigated. Administered concomitantly with acetaminophen, the combination significantly reduced the severity of liver injury. Delayed administration of the combination of drugs, 2 h or 6 h after acetaminophen, also induced a significant decrease in hepatocyte necrosis compared to mice treated with N-acetylcysteine alone. In addition, administration of N-acetylcysteine/glycyrrhizin combination was associated with an improved survival rate compared to mice treated with only N-acetylcysteine.

CONCLUSION

We demonstrate that, compared to N-acetylcysteine alone, co-administration of glycyrrhizin decreases the liver necrosis score and improves survival in a murine model of acetaminophen-induced liver injury. Our study opens a potential new therapeutic pathway in the prevention of acetaminophen hepatotoxicity.

Keywords: Acetaminophen; Acute liver injury; Glycyrrhizin; N-acetylcysteine; N-acetylcysteine/glycyrrhizin combination; Murine model; High mobility group box 1

Core Tip: Acetaminophen overdose is the most common cause of drug-induced liver failure in the developed countries. Substantial progress has been made in understanding the mechanism of hepatocellular injury, but N-acetylcysteine remains the only effective treatment despite its short therapeutic window. We present here our first results on the combination of N-acetylcysteine and glycyrrhizin in a murine model of acetaminophen-induced liver injury. Acetaminophen toxicity was induced by an intraperitoneal dose of 500 mg/kg. Hepatotoxicity was assessed by biochemical and histopathological analyses. Survival rates were also compared. Our results suggest, for the first time, that the combination of N-acetylcysteine and glycyrrhizin may be effective in preventing acetaminophen-induced liver injury in mice.