Abstract
Acetaminophen (APAP)-induced Acute Liver Failure (ALF) is recognized as the most common cause of ALF in Western societies. APAP-induced ALF is characterized by coagulopathy, hepatic encephalopathy, multi-organ failure, and death. MicroRNAs are small, non-coding RNAs that regulate gene expression at the post-transcriptional level. MicroRNA-21 (miR-21) is dynamically expressed in the liver and is involved in the pathophysiology of both acute and chronic liver injury models. We hypothesize that miR-21genetic ablation attenuates hepatotoxicity following acetaminophen intoxication. Eight-week old miR-21knockout (miR21KO) or wild-type (WT) C57BL/6N male mice were injected with acetaminophen (APAP, 300 mg/kg BW) or saline. Mice were sacrificed 6 or 24 h post-injection. MiR21KO mice presented attenuation of liver enzymes ALT, AST, LDH compared with WT mice 24 h post-APAP treatment. Moreover, miR21KO mice had decreased hepatic DNA fragmentation and necrosis than WT mice after 24 h of APAP treatment. APAP-treated miR21KO mice showed increased levels of cell cycle regulators CYCLIN D1 and PCNA, increased autophagy markers expression (Map1LC3a, Sqstm1) and protein (LC3AB II/I, p62), and an attenuation of the APAP-induced hypofibrinolytic state via (PAI-1) compared with WT mice 24 post-APAP treatment. MiR-21 inhibition could be a novel therapeutic approach to mitigate APAP-induced hepatotoxicity and enhance survival during the regenerative phase, particularly to alter regeneration, autophagy, and fibrinolysis. Specifically, miR-21 inhibition could be particularly useful when APAP intoxication is detected at its late stages and the only available therapy is minimally effective.
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Data availability
The datasets generated during and/or analysed during the current study are available from the corresponding author on reasonable request.
Abbreviations
- ALF:
-
Acute liver failure
- ALT:
-
Alanine aminotransferase
- APAP:
-
Acetaminophen
- AU:
-
Arbitrary units
- AST:
-
Aspartate transaminase
- Bcl-2:
-
B-cell lymphoma 2
- Ccnd1:
-
Cyclin D1
- Cyp2e1:
-
Cytochrome P450 2E1
- GSH:
-
Glutathione
- GSSG:
-
Glutathione disulfide
- Hspa1a:
-
Heat shock protein family A member 1A
- JNK:
-
Jun-amino terminal kinase pathway
- Klf6:
-
Activator Kruppel like factor 6
- LDH:
-
Lactate dehydrogenase
- miRNAs:
-
MicroRNAs
- miR-21:
-
MicroRNA-21
- miR21KO:
-
miR-21knockout
- NAC:
-
N-acetylcysteine
- NAPQI:
-
N-acetyl-p-benzoquinoneimine
- PAI-1:
-
Plasminogen activator inhibitor 1
- PCNA:
-
Proliferating cell nuclear antigen
- Pdcd4:
-
Programmed cell death 4
- PINK1:
-
Phosphastase and tensin homolog (PTEN)-induced kinase 1
- Pten:
-
Phosphatase and tensin homolog
- Tnf:
-
Tumor necrosis factor-α
- tPA:
-
Tissue-type plasminogen activator
- TUNEL:
-
Terminal deoxynucleotidyl transferase dUTP nick end labeling assay
- uPA:
-
Urokinase-type plasminogen activator
- WT:
-
Wild-type
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Acknowledgements
We thank the University of Mississippi Medical Center Analytical Assay Core and Imaging Core, and the University of Kansas Medical Center Analytical Core Laboratory for their outstanding service. We thank Dr. Marc E. Rothenberg (Cincinnati Children’s Hospital Medical Center, University of Cincinnati College of Medicine) for generously providing the miR-21 knockout mice. Graphical abstract was created with BioRender.
Funding
Research reported in this publication was supported by the National Institute of General Medical Sciences of the National Institutes of Health under Award Number P20GM121334 (S.R., L.L.Y.C. and D.G.R.) and National Institute of Diabetes and Digestive and Kidney Diseases under Award Number R21DK113500 (D.G.R.). A.M.H. was supported by American Heart Association Predoctoral Fellowship 903804. Research reported in this publication was also supported by the National Institute of General Medical Sciences of the National Institutes of Health under Award P20GM104357 and P20GM121334, P20GM103549, and P30GM118247, and the National Heart, Lung and Blood Institute under Award P01HL51971. The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Institutes of Health.
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AH: conceptualization, formal analysis, investigation, writing—original draft, writing—review and editing, funding acquisition. MS: conceptualization, investigation, writing—review and editing. SR: conceptualization, writing—review and editing, funding acquisition. CA: conceptualization, writing—review and editing. LYC: conceptualization, writing—review and editing, funding acquisition. DR: conceptualization, writing—original draft, writing—review and editing, supervision, funding acquisition.
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Huffman, A.M., Syed, M., Rezq, S. et al. Loss of microRNA-21 protects against acetaminophen-induced hepatotoxicity in mice. Arch Toxicol 97, 1907–1925 (2023). https://doi.org/10.1007/s00204-023-03499-z
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DOI: https://doi.org/10.1007/s00204-023-03499-z