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Assessing Platelet Mitochondrial Dysfunction in a Murine Model of Acute Acetaminophen Toxicity

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Abstract

Introduction

Acetaminophen (APAP) toxicity remains a significant cause of adult and pediatric liver failure in North America and Europe. Previous research has evaluated the impaired mitochondrial function associated with APAP toxicity. The primary aim of this study was to evaluate the effects of APAP toxicity on platelet mitochondrial function using platelet oxygen consumption in a murine model in vivo. Our secondary objectives were to determine the effect of 4-MP on platelet mitochondrial function and hepatic toxicity in the setting of APAP overdose, and to correlate platelet mitochondrial function with other markers of APAP toxicity.

Methods

Male C57Bl/6 mice were randomized to receive APAP (300 or 500 mg/kg) or vehicle followed 90 minutes later by either 4-MP (50 mg/kg) or vehicle via intraperitoneal injection. Mice were euthanized 0, 12, or 24 hours later and platelets isolated from cardiac blood and counted. Platelet oxygen consumption (POC) was determined using a closed-system respirometer. Liver injury was assessed by measuring alanine transferase (ALT) and histological evaluation.

Results

Injection of 500 mg/kg APAP led to increased POC versus pair-matched control (vehicle) (p < 0.001). Administration of 4-MP did not affect POC in control or 300 mg/kg APAP mice. In mice receiving 500 mg/kg APAP and 4-MP, POC decreased significantly compared to mice receiving 500 mg/kg APAP alone (p < 0.01). Serum and histological analysis confirmed APAP-induced hepatic damage in mice receiving 500 mg/kg APAP and these effects blunted by treatment with 4-MP.

Conclusions

Platelet oxygen consumption as a measure of mitochondrial function may be useful as a biomarker of hepatic APAP toxicity in the setting of moderate to severe overdose. Treatment with 4-MP decreases hepatic necrosis and may mitigate the harmful effects of APAP on platelet mitochondrial function detected via POC.

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References

  1. Stravitz RT, Lee WM. Acute liver failure. Lancet. 2019;394(10201):869–81.

    Article  CAS  PubMed  Google Scholar 

  2. Kulkarni S, Perez C, Pichardo C, et al. Use of Pediatric Health Information System database to study the trends in the incidence, management, etiology, and outcomes due to pediatric acute liver failure in the United States from 2008 to 2013. Pediatr Transplant. 2015;19(8):888–95.

    Article  CAS  PubMed  Google Scholar 

  3. Gummin DD, et al. 2019 Annual Report of the American Association of Poison Control Centers’ National Poison Data System (NPDS): 37th Annual Report. Clin Toxicol (Phila). 2020;58(12):1360–541.

    Article  PubMed  Google Scholar 

  4. Budnitz DS, Lovegrove MC, Crosby AE. Emergency department visits for overdoses of acetaminophen-containing products. Am J Prev Med. 2011;40(6):585–92.

    Article  PubMed  Google Scholar 

  5. Akakpo JY, Ramachandran A, Duan L, et al. Delayed treatment with 4-methylpyrazole protects against acetaminophen hepatotoxicity in mice by inhibition of c-Jun n-Terminal Kinase. Toxicol Sci. 2019;170(1):57–68.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  6. Jang DH, Kelly M, Hardy K, Lambert D, Shofer, Eckmann D. A preliminary study in the alterations of mitochondrial respiration in patients with carbon monoxide poisoning measured in blood cells. Clin Toxicol (Phila). 2017;55(6):579–584.

  7. Jang DH, Seeger SC, Grady ME, Shofer FS, Eckmann DM. Mitochondrial dynamics and respiration within cells with increased open pore cytoskeletal meshes. Biol Open. 2017;6(12):1831–9.

    CAS  PubMed  PubMed Central  Google Scholar 

  8. Jang DH, Greenwood JC, Spyres MB, Eckmann DM. Measurement of mitochondrial respiration and motility in acute care: sepsis, trauma, and poisoning. J Intensive Care Med. 2017;32(1):86–94.

    Article  PubMed  Google Scholar 

  9. Zharikov S, Shiva S. Platelet mitochondrial function: from regulation of thrombosis to biomarker of disease. Biochem Soc Trans. 2013;41(1):118–23.

    Article  CAS  PubMed  Google Scholar 

  10. Miyakawa K, Joshi N, Sullivan B, et al. Platelets and protease-activated receptor-4 contribute to acetaminophen-induced liver injury in mice. Blood. 2015;126(15):1835–43.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  11. Lam FW, Rumbaut RE. Platelets mediate acetaminophen hepatotoxicity. Blood. 2017;126(15):1738–9.

    Article  Google Scholar 

  12. Piel S, Chamkha I, Dehlin AK, et al. PLoS One. 2020;15(4):e0231173.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  13. Lauterburg BH, Corcoran GB, Mitchell JR. Mechanism of action of N-acetylcysteine in the protection against the hepatotoxicity of acetaminophen in rats in vivo. J Clin Invest. 1983;71(4):980–91.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  14. Kang AM, Padilla-Jones A, Fisher ES, et al. The effect of 4-methylpyrazole on oxidative metabolism of acetaminophen in human volunteers. J Med Toxicol. 2020;16(2):169–76.

    Article  CAS  PubMed  Google Scholar 

  15. Akakpo JY, Ramachandran A, Kandel SE, et al. 4-Methylpyrazole protects against acetaminophen hepatotoxicity in mice and in primary human hepatocytes. Hum Exp Toxicol. 2018;37(12):1310–22.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  16. Shah K, Beuhler M. Fomepizole as an adjunctive treatment in severe acetaminophen toxicity. Am J Emerg Med. 2020;38(2):410.e5-410.e6.

    Article  PubMed  Google Scholar 

  17. Mossanen JC, Tacke F. Acetaminophen-induced acute liver injury in mice. Lab Anim. 2015;49(1 Suppl):30–6.

    Article  CAS  PubMed  Google Scholar 

  18. Langford D, Bailey A, Chanda M, et al. Coding of facial expressions of pain in the laboratory mouse. Nat Methods. 2010;7(6):447–9.

    Article  CAS  PubMed  Google Scholar 

  19. Ekaney ML, Carrillo-Garcia JC, Gonzalez-Gray G, et al. Platelet aggregation, mitochondrial function and morphology in cold storage: impact of resveratrol and cytochrome c supplementation. Cells. 2022;12(1):166.

    Article  PubMed  PubMed Central  Google Scholar 

  20. Ekaney ML, Grable MA, Powers WF, McKillop IH, Evans SL. Cytochrome c and resveratrol preserve platelet function during cold storage. J Trauma Acute Care Surg. 2017;83(2):271–7.

    Article  CAS  PubMed  Google Scholar 

  21. Thompson K, Nazari SS, Jacobs WC, et al. Use of a crossed high alcohol preferring (cHAP) mouse model with the NIAAA-model of chronic-binge ethanol intake to study liver injury. Alcohol Alcohol. 2017;52(6):629–37.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  22. Sharapova T, Devanarayan V, LeRoy B, et al. Evaluation of miR-122 as a serum biomarker for hepatotoxicity in investigative rat toxicology studies. Vet Pathol. 2016;53(1):211–21.

    Article  CAS  PubMed  Google Scholar 

  23. Liu Y, Li P, Liu L, Zhang Y. The diagnostic role of miR-122 in drug-induced liver injury: A systematic review and meta-analysis. Medicine. 2018;97(49):e13478.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  24. Vatsalya V, Cave MC, Kong M, et al. Keratin 18 is a diagnostic and prognostic factor for acute alcoholic hepatitis. Clin Gastroenterol Hepatol. 2020;18(9):2046–54.

    Article  CAS  PubMed  Google Scholar 

  25. Umbaugh DS, Jaeschke H. Biomarkers of drug-induced liver injury: a mechanistic perspective through acetaminophen hepatotoxicity. Expert Rev Gastroenterol Hepatol. 2021;15(4):363–75.

    Article  CAS  PubMed  Google Scholar 

  26. James LP, Letzig L, Simpson PM, et al. Pharmacokinetics of acetaminophen-protein adducts in adults with acetaminophen overdose and acute liver failure. Drug Metab Dispos. 2009;37(8):1779–84.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  27. Liu J, Jiang M, Jin Q, et al. Modulation of HMGB1 release in APAP-induced liver injury: a possible strategy of Chikusetsusaponin V targeting NETs formation. Front Pharmacol. 2021;12:723881.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  28. Sjövall F, Morota S, Persson J, et al. Patients with sepsis exhibit increased mitochondrial respiratory capacity in peripheral blood immune cells. Crit Care. 2013;17:R152.

    Article  PubMed  PubMed Central  Google Scholar 

  29. Decision Tree. [Mouse blood sampling decision tree]. Available from: https://www.nc3rs.org.uk/mouse-decision-tree-blood-sampling. Accessed 1 Dec 2019.

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Funding

2020 Medical Toxicology Foundation (MTF) Innovative Research and Teaching Award.

Author information

Authors and Affiliations

  1. Department of Emergency Medicine, Atrium Health’s Carolinas Medical Center, 1000 Blythe Blvd., Medical Education Building 3rd Floor, Charlotte, NC, 28203, USA

    Carolyn Fox, Michael Runyon & Christine M. Murphy

  2. Department of Surgery, Atrium Health’s Carolinas Medical Center, 1000 Blythe Blvd., Cannon Research Building, Charlotte, NC, 28203, USA

    Michael L. Ekaney & Iain H. McKillop

  3. Center for Outcomes Research and Evaluation (CORE), 1300 Scott Ave, Office 124, Charlotte, NC, 28204, USA

    Hieu M. Nguyen

  4. Department of Data Science, University of Mississippi Medical Center, 2500 N State St, Jackson, MS, 39216, USA

    Philip J. Turk

Authors
  1. Carolyn Fox
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  2. Michael L. Ekaney
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  3. Michael Runyon
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  4. Hieu M. Nguyen
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  5. Philip J. Turk
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  6. Iain H. McKillop
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  7. Christine M. Murphy
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Contributions

All authors contributed to the study's conception and design. Material preparation was performed by Carolyn Fox, Christine Murphy, and Michael Ekaney. Data collection was performed by Carolyn Fox, Christine Murphy, Michael Ekaney, and Iain McKillop. Data analysis was performed by all authors. Philip Turk and Hieu Nguyen performed the POC analysis, while the other analysis was performed by the remaining authors. All authors contributed to the original draft of the manuscript, and all authors participated in review and editing of the manuscript. All authors read and approved the final manuscript.

Corresponding author

Correspondence to Christine M. Murphy.

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Ethical Approval

All institutional and national guidelines for the care and use of laboratory animals were followed.

Conflict of Interest

The authors declare that they have no conflicts of interest.

Additional information

Supervising Editor: Anselm Wong, MBBS DipTox, PhD

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Previous Presentation: Data in this study were previously presented at the American College of Medical Toxicology (ACMT) Annual Scientific Meeting, April 2021.

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Fox, C., Ekaney, M.L., Runyon, M. et al. Assessing Platelet Mitochondrial Dysfunction in a Murine Model of Acute Acetaminophen Toxicity. J. Med. Toxicol. 19, 341–351 (2023). https://doi.org/10.1007/s13181-023-00964-0

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  • DOI: https://doi.org/10.1007/s13181-023-00964-0

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