Hepatotoxic effect of acetaminophen on liver tissue of Sprague-Dawley rat fetuses: Histopathological, biochemical, and implications for hepatocellular carcinoma risk
DOI:
https://doi.org/10.37800/RM.3.2026.803Keywords:
acetaminophen, paracetamol, hepatotoxicity, fetal development, pregnancy, hepatocellular carcinoma, oxidative stress, developmental toxicityAbstract
Relevance: Acetaminophen (paracetamol) is one of the most commonly used analgesic-antipyretic agents during pregnancy. Prenatal hepatotoxic exposures may influence fetal liver development and affect susceptibility to hepatocellular carcinoma in offspring.
The study aimed to describe the dose-dependent hepatotoxic effects of prenatal acetaminophen exposure on fetal liver tissues in Sprague-Dawley rats and assess their relevance to risk pathways for human hepatocellular carcinoma.
Materials and Methods: On gestation days (GD) 6-15, pregnant Sprague-Dawley rats were randomly assigned to four groups: Control (normal saline), Low Dose (LDG; 200 mg/kg), High Dose (HDG; 400 mg/kg), and Recovery (RCG; 400 mg/kg followed by withdrawal). Serum liver enzymes, hepatic oxidative stress markers, molecular markers, and histopathological changes were assessed in fetal livers at gestational day 20.
Results: Hepatotoxicity was dose-dependent. The HDG group showed markedly elevated alanine aminotransferase (ALT: 78.6 ± 5.8 vs. 28.5 ± 2.1 U/L), aspartate aminotransferase (AST: 98.4 ± 7.2 vs. 35.2 ± 2.8 U/L), and alkaline phosphatase (ALP: 125.6 ± 8.5 vs. 68.5 ± 4.2 U/L; all p < 0.001). Malondialdehyde (MDA) levels increased 3.8-fold, while glutathione (GSH) levels were reduced by 66%, with significant decreases in superoxide dismutase (SOD) and catalase (CAT) activity. Histopathology revealed a spectrum of changes ranging from LDG to extensive HDG. Cytochrome P450 2E1 (CYP2E1) expression was upregulated 4.9-fold in the HDG group; p53 and nuclear factor kappa-B p65 (NF-κB p65) were markedly overexpressed. The Bax/Bcl-2 ratio and cleaved caspase-3 levels showed dose-dependent pro-apoptotic activation. The RCG group showed partial recovery.
Conclusion: Acetaminophen exerts dose-dependent fetal hepatotoxicity, mediated by oxidative stress (glutathione depletion, lipid peroxidation, and suppression of antioxidant enzymes) and the activation of molecular pathways relevant to hepatocellular carcinoma. These findings highlight the need for caution regarding acetaminophen administration during pregnancy and support further longitudinal investigation of its potential role in oncogenic programming.
References
1. Yoon E, Babar A, Choudhary M, Kutner M, Pyrsopoulos N. Acetaminophen-induced hepatotoxicity: a comprehensive update. J Clin Transl Hepatol. 2016;4(2):131-142.
https://doi.org/10.14218/JCTH.2015.00052
2. Zafeiri A, Mitchell RT, Hay DC, Fowler PA. Over-the-counter analgesics during pregnancy: a comprehensive review of global prevalence and offspring safety. Hum Reprod Update. 2021;27(1):67-95.
https://doi.org/10.1093/humupd/dmaa042
3. Taagaard M, Rode L, de Wolff MG, Damm P, Hagen CP, Fisher MB, Hegaard HK, Rom AL. Paracetamol use prior to and in early pregnancy: prevalence and patterns among women with and without chronic medical diseases. Br J Clin Pharmacol. 2023;89(8):2582-2591.
https://doi.org/10.1111/bcp.15732
4. Bauer AZ, Kriebel D, Herbert MR, Bornehag CG, Swan SH. Prenatal paracetamol exposure and child neurodevelopment: a review. Horm Behav. 2018;101:125-147.
https://doi.org/10.1016/j.yhbeh.2018.01.003
5. Bauer AZ, Swan SH, Kriebel D, Liew Z, Taylor HS, Bornehag CG, Andrade AM, Olsen J, Jensen RH, Mitchell RT, Skakkebaek NE, Jégou B, Kristensen DM. Paracetamol use during pregnancy — a call for precautionary action. Nat Rev Endocrinol. 2021;17:757-766.
https://doi.org/10.1038/s41574-021-00553-7
6. Lu Z, Guo Y, Xu D, Xiao H, Dai Y, Liu K, Chen L, Wang H. Developmental toxicity and programming alterations of multiple organs in offspring induced by medication during pregnancy. Acta Pharm Sin B. 2023;13(2):460-477.
https://doi.org/10.1016/j.apsb.2022.05.029
7. Wu K, Lu W, Yan X. Potential adverse actions of prenatal exposure of acetaminophen to offspring. Front Pharmacol. 2023;14:1094435.
https://doi.org/10.3389/fphar.2023.1094435
8. Xu M, Wang R, Du B, Zhang Y, Feng X. Association of acetaminophen use with perinatal outcomes among pregnant women: a retrospective cohort study with propensity score matching. BMC Pregnancy Childbirth. 2024;24(1):268.
https://doi.org/10.1186/s12884-024-06480-5
9. Liu Y, Liu Y, Chen S, Kong Z, Guo Y, Wang H. Prenatal exposure to acetaminophen at different doses, courses and time causes testicular dysplasia in offspring mice and its mechanism. Chemosphere. 2023;345:140496.
https://doi.org/10.1016/j.chemosphere.2023.140496
10. Kamath P, Kamath A, Ullal SD. Liver injury associated with drug intake during pregnancy. World J Hepatol. 2021;13(7):747-762.
https://doi.org/10.4254/wjh.v13.i7.747
11. Al-Rekabi SAM, Talib Abed W, Al-Shemary NNA. Impact of aspirin suppositories on fertility in female mice. Indian J Forensic Med Toxicol. 2019;13(4):1505-1509.
https://doi.org/10.5958/0973-9130.2019.00515.2
12. Al-Rikabi SAM, Al-Shemary NNA. The effect of letrozole treatment on fertility and embryo parameters in female mice: histological and hormonal study. J Pak Med Assoc. 2023;73(9):S150-S154.
https://doi.org/10.47391/JPMA.IQ-31
13. Abid Al-Shemary NN, Mousa SA, Muslim ZZ. Histological and hormonal study of the effect of aqueous extract of Ocimum gratissimum on female reproductive system in albino mice. J Pharm Sci Res. 2018;10(4):765-767.
14. Hussain MJ, Sarboot AK, Hussein EF. Histological study of celery aqueous seed extract (Apium graveolens L.) on ovaries in adult female rats. Plant Arch. 2018;18(2):2717-2722.
15. Ramachandran A, Jaeschke H. Oxidant stress and acetaminophen hepatotoxicity: mechanism-based drug development. Antioxid Redox Signal. 2021;35(9):718-733.
https://doi.org/10.1089/ars.2021.0102
16. Luo G, Huang L, Zhang Z. The molecular mechanisms of acetaminophen-induced hepatotoxicity and its potential therapeutic targets. Exp Biol Med (Maywood). 2023;248(5):412-424.
https://doi.org/10.1177/15353702221147563
17. Andrade RJ, Chalasani N, Björnsson ES, Suzuki A, Kullak-Ublick GA, Watkins PB, Devarbhavi H, Merz M, Lucena MI, Kaplowitz N, Aithal GP. Drug-induced liver injury. Nat Rev Dis Primers. 2019;5(1):58.
https://doi.org/10.1038/s41572-019-0105-0
18. Jaeschke H, Ramachandran A. Acetaminophen hepatotoxicity: paradigm for understanding mechanisms of drug-induced liver injury. Annu Rev Pathol. 2024;19:453-478.
https://doi.org/10.1146/annurev-pathmechdis-051122-094016
19. Cai X, Cai H, Wang J, Yang Q, Guan J, Deng J, Chen Z. Molecular pathogenesis of acetaminophen-induced liver injury and its treatment options. J Zhejiang Univ Sci B. 2022;23(4):265-285.
https://doi.org/10.1631/jzus.B2100977
20. Yan M, Huo Y, Yin S, Hu H. Mechanisms of acetaminophen-induced liver injury and its implications for therapeutic interventions. Redox Biol. 2018;17:274-283.
https://doi.org/10.1016/j.redox.2018.04.019
21. Li Y, Yu Y, Yang L, Wang R. Insights into the role of oxidative stress in hepatocellular carcinoma development. Front Biosci (Landmark Ed). 2023;28(11):286.
https://doi.org/10.31083/j.fbl2811286
22. Bruix J, Reig M, Sherman M. Evidence-based diagnosis, staging, and treatment of patients with hepatocellular carcinoma. Gastroenterology. 2016;150(4):835-853.
https://doi.org/10.1053/j.gastro.2015.12.041
23. Chapron BD, Chapron A, Leeder JS. Recent advances in the ontogeny of drug disposition. Br J Clin Pharmacol. 2022;88(10):4267-4284. Epub 2021.
https://doi.org/10.1111/bcp.14821
24. van Groen BD, Nicolaï J, Kuik AC, Van Cruchten S, van Peer E, Smits A, Schmidt S, de Wildt SN, Allegaert K, De Schaepdrijver L, Annaert P, Badée J. Ontogeny of hepatic transporters and drug-metabolizing enzymes in humans and in nonclinical species. Pharmacol Rev. 2021;73(2):597-678.
https://doi.org/10.1124/pharmrev.120.000071
25. Sevrioukova IF. Structural basis for the diminished ligand binding and catalytic ability of human fetal-specific CYP3A7. Int J Mol Sci. 2021;22(11):5831.
https://doi.org/10.3390/ijms22115831
26. Lewis K, Yoshimoto M, Takebe T. Fetal liver hematopoiesis: from development to delivery. Stem Cell Res Ther. 2021;12(1):139.
https://doi.org/10.1186/s13287-021-02189-w
27. Nobile S, Di Sipio Morgia C, Vento G. Perinatal origins of adult disease and opportunities for health promotion: a narrative review. J Pers Med. 2022;12(2):157.
https://doi.org/10.3390/jpm12020157
28. Lapehn S, Paquette AG. The placental epigenome as a molecular link between prenatal exposures and fetal health outcomes through the DOHaD hypothesis. Curr Environ Health Rep. 2022;9(3):490-501.
https://doi.org/10.1007/s40572-022-00354-8
29. Wu T, Zhou K, Hua Y, Zhang W, Li Y. The molecular mechanisms in prenatal drug exposure-induced fetal programmed adult cardiovascular disease. Front Pharmacol. 2023;14:1164487.
https://doi.org/10.3389/fphar.2023.1164487
30. Mian P, van den Anker JN, van Calsteren K, Annaert P, Tibboel D, Pfister M, Allegaert K, Dallmann A. Physiologically based pharmacokinetic modeling to characterize acetaminophen pharmacokinetics and N-acetyl-p-benzoquinone imine (NAPQI) formation in non-pregnant and pregnant women. Clin Pharmacokinet. 2020;59(1):97-110. Epub 2019.
https://doi.org/10.1007/s40262-019-00799-5
31. Castro CT, Gama RS, Pereira M, Oliveira MG, Dal-Pizzol TS, Barreto ML, Santos DB. Effect of acetaminophen use during pregnancy on adverse pregnancy outcomes: a systematic review and meta-analysis. Expert Opin Drug Saf. 2022;21(2):241-251.
https://doi.org/10.1080/14740338.2022.2020246
32. Michalopoulos GK, Bhushan B. Liver regeneration: biological and pathological mechanisms and implications. Nat Rev Gastroenterol Hepatol. 2021;18(1):40-55.
https://doi.org/10.1038/s41575-020-0342-4
33. Campana L, Esser H, Huch M, Forbes S. Liver regeneration and inflammation: from fundamental science to clinical applications. Nat Rev Mol Cell Biol. 2021;22(9):608-624.
https://doi.org/10.1038/s41580-021-00373-7
34. Jaeschke H, Ramachandran A. Central mechanisms of acetaminophen hepatotoxicity: mitochondrial dysfunction by protein adducts and oxidant stress. Drug Metab Dispos. 2024;52(8):712-721.
https://doi.org/10.1124/dmd.123.001279
35. Yang T, Wang H, Wang X, Li J, Jiang L. The dual role of innate immune response in acetaminophen-induced liver injury. Biology (Basel). 2022;11(7):1057.
https://doi.org/10.3390/biology11071057
36. Jaeschke H, Ramachandran A. Mechanisms and pathophysiological significance of sterile inflammation during acetaminophen hepatotoxicity. Food Chem Toxicol. 2020;138:111240.
https://doi.org/10.1016/j.fct.2020.111240
37. Al-Ogaili AS, Hameed SS, Noori N. LPS-induced NLRP3 gene-expression in chicken. Open Vet J. 2022;12(2):197-203.
https://doi.org/10.5455/OVJ.2022.v12.i2.7
38. Licata A, Minissale MG, Stankevičiūtė S, Sanabria-Cabrera J, Lucena MI, Andrade RJ, Almasio PL. N-acetylcysteine for preventing acetaminophen-induced liver injury: a comprehensive review. Front Pharmacol. 2022;13:828565.
https://doi.org/10.3389/fphar.2022.828565
39. Ramachandran A, Jaeschke H. A mitochondrial journey through acetaminophen hepatotoxicity. Food Chem Toxicol. 2020;140:111282.
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