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Digestive Diseases and Sciences

, Volume 63, Issue 12, pp 3382–3397 | Cite as

3-Aminobenzamide Prevents Concanavalin A-Induced Acute Hepatitis by an Anti-inflammatory and Anti-oxidative Mechanism

  • Joram WardiEmail author
  • Orna Ernst
  • Anna Lilja
  • Hussein Aeed
  • Sebastián Katz
  • Idan Ben-Nachum
  • Iris Ben-Dror
  • Dolev Katz
  • Olga Bernadsky
  • Rajendar Kandhikonda
  • Yona Avni
  • Iain D. C. Fraser
  • Roy Weinstain
  • Alexander BiroEmail author
  • Tsaffrir ZorEmail author
Original Article

Abstract

Background and Aims

Concanavalin A is known to activate T cells and to cause liver injury and hepatitis, mediated in part by secretion of TNFα from macrophages. Poly(ADP-ribose) polymerase-1 (PARP-1) inhibitors have been shown to prevent tissue damage in various animal models of inflammation. The objectives of this study were to evaluate the efficacy and mechanism of the PARP-1 inhibitor 3-aminobenzamide (3-AB) in preventing concanavalin A-induced liver damage.

Methods

We tested the in vivo effects of 3-AB on concanavalin A-treated mice, its effects on lipopolysaccharide (LPS)-stimulated macrophages in culture, and its ability to act as a scavenger in in vitro assays.

Results

3-AB markedly reduced inflammation, oxidative stress, and liver tissue damage in concanavalin A-treated mice. In LPS-stimulated RAW264.7 macrophages, 3-AB inhibited NFκB transcriptional activity and subsequent expression of TNFα and iNOS and blocked NO production. In vitro, 3-AB acted as a hydrogen peroxide scavenger. The ROS scavenger N-acetylcysteine (NAC) and the ROS formation inhibitor diphenyleneiodonium (DPI) also inhibited TNFα expression in stimulated macrophages, but unlike 3-AB, NAC and DPI were unable to abolish NFκB activity. PARP-1 knockout failed to affect NFκB and TNFα suppression by 3-AB in stimulated macrophages.

Conclusions

Our results suggest that 3-AB has a therapeutic effect on concanavalin A-induced liver injury by inhibiting expression of the key pro-inflammatory cytokine TNFα, via PARP-1-independent NFκB suppression and via an NFκB-independent anti-oxidative mechanism.

Keywords

Liver failure Inflammation Macrophages TNFα Reactive oxygen species NFκB 

Abbreviations

3-AB

3-Aminobenzamide

ALT

Alanine aminotransferase

AST

Aspartate aminotransferase

ConA

Concanavalin A

DPI

Diphenyleneiodonium

HRP

Horseradish peroxidase

IFNγ

Interferon γ

LPS

Lipopolysaccharide

MDH

Malate dehydrogenase

NAC

N-acetylcysteine

NFκB

Nuclear factor kappa B

NO

Nitric oxide

Nox

NADPH oxidase

PARP-1

Poly(ADP-ribose) polymerase-1

PF2

Peroxyfluor-2

ROS

Reactive oxygen species

SOD

Superoxide dismutase

TNFα

Tumor necrosis factor α

Notes

Acknowledgments

We are grateful to Dr. C. Daniels and M. Athamna for critical reading of the manuscript, to Dr. M. Cohen-Armon for various reagents and helpful discussions, to Dr. E. Pick and Dr. E. Bechor for the help with the superoxide assay and insightful discussions; to Dr. D. Shabat, N. Hananya and O. Green for help with fluorescence measurement; to Dr. C. Yu (Xiamen University, Xiamen, Fujian, China) for the TNFα promoter luciferase plasmid; to Dr. M. Montminy (Salk Institute, La-Jolla, CA) for the CRE-luciferase plasmid; to Dr. P. Angel (German Cancer Research Center, Heidelberg, Germany) for the AP-1-luciferase plasmid; to Dr. Ariel Munitz (TAU, Israel) for the iNOS antibody; and to Dr. Y. Ebenstein (TAU, Israel) for the EL-4 cell line.

Author’s contribution

JW, AB and TZ conceived and designed the study; OE, AL, HA, SK, IBN, IBD, DK, and KR performed the experiments; JW, OE, AL, HA, SK, DK, OB, KR, IF, RW, AB, and TZ analyzed the data; JW, OE, AB, and TZ wrote the manuscript.

Funding

This work was supported by the United States – Israel Binational Science Foundation [Grant 2011360 to TZ]. IF is supported by the Intramural Research Program of NIAID, NIH.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

Ethics approval and consent to participate

All animal experiments were performed in accordance with the guidelines of the Care and Use of Laboratory Animals and have been approved by the research ethics committee at Wolfson medical center.

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© Springer Science+Business Media, LLC, part of Springer Nature 2018

Authors and Affiliations

  1. 1.Department of GastroenterologyE. Wolfson Medical CenterHolonIsrael
  2. 2.Department of Biochemistry and Molecular Biology, Life Sciences FacultyTel-Aviv UniversityTel-AvivIsrael
  3. 3.Signaling Systems Section, Laboratory of Immune System Biology, National Institute of Allergy and Infectious DiseasesNational Institutes of HealthBethesdaUSA
  4. 4.Department of PathologyE. Wolfson Medical CenterHolonIsrael
  5. 5.Department of Molecular Biology and Ecology of Plants, Life Sciences FacultyTel-Aviv UniversityTel-AvivIsrael
  6. 6.Institute of NephrologyE. Wolfson Medical CenterHolonIsrael

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