Sıçanlarda Metotreksat ile İndüklenen Karaciğer Toksisitesinde Polidatinin Koruyucu Etkileri; Çoklu Yolak Analizi

Abstract

Bu çalışma, polidatinin metotreksat (MTX) ile indüklenen hepatotoksisiteye karşı koruyucu etkilerini sıçanlarda değerlendirmiştir. Toplam 35 Wistar albino sıçan şu gruplara ayrılmıştır: Kontrol, yalnızca polidatin (100 mg/kg), yalnızca MTX (20 mg/kg), MTX + polidatin (50 mg/kg) ve MTX + polidatin (100 mg/kg). Tedaviler 7 gün süreyle uygulanmıştır. MTX uygulaması, serum AST/ALT düzeylerinde, hepatik malondialdehit (MDA) düzeylerinde ve proinflamatuvar/apoptotik belirteçlerde (NF-κB, TNF-α, Bax, Kaspaz-3 mRNA) anlamlı artışa neden olurken; antiapoptotik Bcl-2 ve doku onarımı ile ilişkili sinyaller (Wnt3 mRNA) azalmıştır. Polidatin ile birlikte uygulama bu etkileri doza bağımlı olarak tersine çevirmiştir. 100 mg/kg doz, AST/ALT ve MDA düzeylerini 50 mg/kg doza kıyasla daha etkili şekilde normalize etmiştir (p<0,05), ancak her iki doz da TNF-α, Bax ve Kaspaz-3 düzeylerini benzer şekilde baskılamıştır. Ayrıca, 100 mg/kg polidatin uygulaması, düşük doza kıyasla Bcl-2 ve Wnt3 ekspresyonunu anlamlı olarak artırmıştır (p<0,05). Antioksidan savunma sistemleri (SOD, CAT, GPx, GSH) de doza bağımlı şekilde yeniden düzelmiştir. Histopatolojik inceleme, MTX gruplarında belirgin inflamasyon ve hücresel hasarı doğrularken, polidatin uygulanan gruplarda karaciğer mimarisinin normale yakın düzeyde korunduğunu göstermiştir. Elde edilen bulgular, polidatinin oksidatif stresi azaltma, inflamasyonu ve apoptozu baskılama ve rejenerasyonu artırma yoluyla MTX’e bağlı karaciğer hasarına karşı çok hedefli koruyucu etki gösterdiğini ve bu nedenle adjuvan tedavi açısından potansiyel taşıdığını ortaya koymaktadır.

Keywords

• Polidatin
• Apoptozis
• Methotreksat
• Okidadatif stres
• Hepatotoksisite

Ethical Statement

Tüm hayvanları içeren deneysel prosedürler, laboratuvar hayvanlarının bakımı ve kullanımı için kurumsal ve ulusal yönergelere uygun olarak yürütülmüştür.

Protective Effects of Polydatin in Methotrexate-Induced Liver Toxicity in Rats; Multi-Pathway Analysis

Abstract

This study evaluated polydatin's protective effects against methotrexate (MTX)-induced hepatotoxicity in rats. Thirty-five Wistar albino rats were divided into (n=7): Control, Polydatin-only (100 mg/kg), MTX-only (20 mg/kg), MTX + Polydatin (50 mg/kg), and MTX + Polydatin (100 mg/kg). Treatments lasted 7 days. MTX administration significantly increased serum AST/ALT levels, hepatic malondialdehyde (MDA), and pro-inflammatory/apoptotic markers (NF-κB, TNF-α, Bax, Caspase-3 mRNA), while decreasing anti-apoptotic Bcl-2 and tissue-repair signals (Wnt3 mRNA). Co-treatment with polydatin dose-dependently reversed these effects: the 100 mg/kg dose notably normalized AST/ALT and MDA levels more effectively than 50 mg/kg (p<0.05), while both doses comparably suppressed TNF-α, Bax, and Caspase-3. Polydatin (100 mg/kg) significantly elevated Bcl-2 and Wnt3 expression versus lower-dose treatment (p<0.05). Antioxidant defenses (SOD, CAT, GPx, GSH) were also restored in a dose-responsive manner. Histopathology confirmed severe inflammation and cellular damage in MTX groups, whereas polydatin groups showed near-normal liver architecture. Findings demonstrate polydatin’s multi-target protection against MTX induced liver injury through mitigating oxidative stress, inflammation, apoptosis, and enhancing regeneration, supporting its adjunctive therapeutic potential.

Keywords

• Methotrexate hepatotoxicity
• Polydatin
• Oxidative stress
• Apoptosis

Ethical Statement

All experimental procedures involving animals were conducted in accordance with institutional and national guidelines for the care and use of laboratory animals.

References

1. Alqarni A, Zeidler M. “How does methotrexate work?.” Biochemical Society Transactions 48.2 (2020): 559-567.
2. Chan E, Cronstein B. Methotrexate—how does it really work? Nat Rev Rheumatol. 2010 Mar 1;6:175–8. doi:10.1038/nrrheum.2010.5
3. Cronstein B. Low-Dose Methotrexate: A Mainstay in the Treatment of Rheumatoid Arthritis. Pharmacol Rev. 2005 Jun 1;57:163–72. doi:10.1124/pr.57.2.3
4. Bedoui Y, Guillot X, Selambarom J, Guiraud P, Giry C, Jaffar-Bandjee M, at al. Methotrexate an Old Drug with New Tricks. Int J Mol Sci [Internet]. 2019 Oct 1;20.
5. Cronstein B, Aune T. Methotrexate and its mechanisms of action in inflammatory arthritis. Nat Rev Rheumatol. 2020 Feb 17;16:145–54. doi:10.1038/s41584-020-0373-9
6. Cronstein B, Naime D, Ostad E. The antiinflammatory mechanism of methotrexate. Increased adenosine release at inflamed sites diminishes leukocyte accumulation in an in vivo model of inflammation. J Clin Invest. 1993 Dec 1;92 6:2675–82. doi:10.1172/JCI116884
7. Maksimovic V, Pavlović-Popović Z, Vukmirović S, Cvejić J, Mooranian A, Al‐Salami H, at al. Molecular mechanism of action and pharmacokinetic properties of methotrexate. Mol Biol Rep. 2020 May 15;47:4699–708.
8. Kandemir FM, Kucukler S, Caglayan C, Gur C, Batil AA, Gülçin İ. Therapeutic effects of silymarin and naringin on methotrexate‐induced nephrotoxicity in rats: Biochemical evaluation of anti‐inflammatory, antiapoptotic, and antiautophagic properties. J Food Biochem. 2017;41(5):e12398.

9. Li Z, Chen X, Liu G, Li J, Zhang J, Cao Y, at al. Antioxidant activity and mechanism of resveratrol and polydatin isolated from mulberry (Morus alba L.). Molecules. 2021;26(24):1–18.
10. Deminice R, Da Silva RP, Lamarre SG, Kelly KB, Jacobs RL, Brosnan ME, at al. Betaine supplementation prevents fatty liver induced by a high-fat diet: Effects on one-carbon metabolism. Amino Acids. 2015 Jan 11;47(4):839–46.
11. Khan ZA, Tripathi R, Mishra B. Methotrexate: A detailed review on drug delivery and clinical aspects. Expert Opin Drug Deliv. 2012 Feb;9(2):151–69. doi:10.1517/17425247.2012.642362 PubMed PMID: 22251428.
12. Cheng K, Niu J, Zheng X, Qiao Y, Zhang J, Guo R, Dong G, Song Z, Huang J, Wang J. Aflatoxin-B1-exposure-induced hepatic injury could be alleviated by polydatin through reducing oxidative stress, inhibiting inflammation and improving mitophagy. Toxics. 2023;11(4):309.
13. Stochino Loi E, Pontis A, Cofelice V, Pirarba S, Fais MF, Daniilidis A, Melis I, Paoletti AM, Angioni S. Effect of ultramicronized-palmitoylethanolamide and co-micronized palmitoylethanolamide/polydatin on chronic pelvic pain and quality of life in endometriosis patients: an open-label pilot study. Int J Womens Health. 2019;443–9.
14. Tang D, Zhang Q, Duan H, Ye X, Liu J, Peng W, Wu C. Polydatin: a critical promising natural agent for liver protection via antioxidative stress. Oxid Med Cell Longev. 2022;2022(1):9218738.
15. Ye G, Zhao Y, Zhu J, Zhang Z, Wang Q, Jiang X, Wang Z. Synergistic Effect of Polydatin and Polygonatum sibiricum Polysaccharides in Combating Atherosclerosis via Suppressing TLR4‐Mediated NF‐κB Activation in ApoE‐Deficient Mice. Evidence‐Based Complement Altern Med. 2022;2022(1):3885153.
16. Ahmad P, Alvi SS, Iqbal D, Khan MS. Insights into pharmacological mechanisms of polydatin in targeting risk factors-mediated atherosclerosis. Life Sci. 2020;254:117756.
17. Placer ZA, Cushman LL, Johnson BC. Estimation of product of lipid peroxidation (malonyl dialdehyde) in biochemical systems. Anal Biochem. 1966;16(2):359–64.
18. Sedlak J, Lindsay RH. Estimation of total, protein-bound, and nonprotein sulfhydryl groups in tissue with Ellman’s reagent. Anal Biochem. 1968;25:192–205.
19. Lowry OH, Rosebrough NJ, Farr AL, Randall RJ. Protein measurement with the Folin phenol reagent. [Internet]. 1951 [cited 2025 Apr 23]. Available from: https://www.cabidigitallibrary.org/doi/full/10.5555/19511404458
20. Livak KJ, Schmittgen TD. Analysis of relative gene expression data using real-time quantitative PCR and the 2- ΔΔCT method. methods. 2001;25(4):402–8.
21. Segal R, Yaron M, Tartakovsky B. Methotrexate: mechanism of action in rheumatoid arthritis. Semin Arthritis Rheum. 1990 Dec 1;20 3:190–200. doi:10.1016/0049-0172(90)90060-S
22. Koźmiński P, Halik P, Chesori R, Gniazdowska E. Overview of Dual-Acting Drug Methotrexate in Different Neurological Diseases, Autoimmune Pathologies and Cancers. Int J Mol Sci. 2020 May 1;21. doi:10.3390/ijms21103483
23. El-Esawy FM, Ahmed IA, El-Fallah AA, Salem RM. Methotrexate mechanism of action in plaque psoriasis: something new in the old view. J Clin Aesthetic Dermatol. 2022;15(8):42.
24. Zhao Z, Hua Z, Luo X, Li Y, Yu L, Li M, Lu C, Zhao T, Liu Y. Application and pharmacological mechanism of methotrexate in rheumatoid arthritis. Biomed Pharmacother Biomedecine Pharmacother. 2022 Jun 1;150:113074. doi:10.1016/j.biopha.2022.113074
25. Aboubakr EM, Ibrahim ARN, Ali FEM, Mourad AAE, Ahmad AM, Hofni A. Fasudil Ameliorates Methotrexate-Induced Hepatotoxicity by Modulation of Redox-Sensitive Signals. Pharmaceuticals. 2022 Nov 19;15(11):1436. doi:10.3390/ph15111436
26. Friedman B, Cronstein B. Methotrexate mechanism in treatment of rheumatoid arthritis. Joint Bone Spine. 2019 May;86(3):301–7. doi:10.1016/j.jbspin.2018.07.004
27. Kalantari E, Zolbanin NM, Ghasemnejad-Berenji M. Protective effects of empagliflozin on methotrexate induced hepatotoxicity in rats. Biomed Pharmacother. 2024 Jan;170:115953. doi:10.1016/j.biopha.2023.115953
28. Erişir M, Benzer F, Kandemir FM. Changes in the rate of lipid peroxidation in plasma and selected blood antioxidants before and during pregnancy in ewes. Acta Vet Brno. 2009;78(2):237–42.
29. Kandemir O, Kucukler S, Comakli S, Gur C, Ileriturk M. Docetaxel-induced liver and kidney toxicity in rats can be alleviated by suppressing oxidative stress, endoplasmic reticulum stress, inflammation, apoptosis and autophagy signaling pathways after Silymarin treatment. Food Chem Toxicol. 2025;196:115202.
30. Khafaga AF, El-Sayed YS. Spirulina ameliorates methotrexate hepatotoxicity via antioxidant, immune stimulation, and proinflammatory cytokines and apoptotic proteins modulation. Life Sci. 2018 Mar;196:9–17. doi:10.1016/j.lfs.2018.01.010
31. Kalantari H, Asadmasjedi N, Abyaz MR, Mahdavinia M, Mohammadtaghvaei N. Protective effect of inulin on methotrexate- induced liver toxicity in mice. Biomed Pharmacother. 2019 Feb;110:943–50. doi:10.1016/j.biopha.2018.11.144
32. El Masri AER, Tobler C, Willemijn B, Von Bueren AO, Ansari M, Samer CF. Case report: Hepatotoxicity and nephrotoxicity induced by methotrexate in a paediatric patient, what is the role of precision medicine in 2023? Front Pharmacol. 2023 May 2;14:1130548. doi:10.3389/fphar.2023.1130548
33. Eriten B, Caglayan C, Gür C, Küçükler S, Diril H. Hepatoprotective effects of zingerone on sodium arsenite-induced hepatotoxicity in rats: Modulating the levels of caspase-3/Bax/Bcl-2, NLRP3/NF-κB/TNF-α and ATF6/IRE1/PERK/GRP78 signaling pathways. Biochem Biophys Res Commun. 2024 Sep 17;725:150258. doi:10.1016/j.bbrc.2024.150258
34. Erzincan R, Caglayan C, Kandemir FM, İzol E, Gür C, İleritürk M. Hepatoprotective Effects of Royal Jelly Against Vincristine-Induced Hepatotoxicity in Rats: A Biochemical and Molecular Study. Life. 2025 Mar;15(3):459. doi:10.3390/life15030459
35. Kandemir O, Kucukler S, Comakli S, Gur C, İleriturk M. Docetaxel-induced liver and kidney toxicity in rats can be alleviated by suppressing oxidative stress, endoplasmic reticulum stress, inflammation, apoptosis and autophagy signaling pathways after Silymarin treatment. Food Chem Toxicol. 2025 Feb 1;196:115202. doi:10.1016/j.fct.2024.115202
36. Caglayan C, Ekinci İ, Gur C, Ayna A, Bayav İ, Kandemir FM. Protective Effects of Chrysin Against Diclofenac-Induced Nephrotoxicity in Rats via Attenuation of Oxidative Stress, Apoptosis and Endoplasmic Reticulum Stress. J Biochem Mol Toxicol. 2025;39(6):e70373. doi:10.1002/jbt.70373
37. Gezer A, Üstündağ H, Karadağ Sarı E, Bedir G, Gür C, Mendil AS, Duysak L. β-carotene protects against α-amanitin nephrotoxicity via modulation of oxidative, autophagic, nitric oxide signaling, and polyol pathways in rat kidneys. Food Chem Toxicol. 2024 Nov 1;193:115040. doi:10.1016/j.fct.2024.115040
38. Cakmak F, Kucukler S, Gur C, Ileriturk M, Gul M, Varisli B. Cardiotoxicity caused by acrylamide in rats can be alleviated as a result of suppression of oxidative stress, endoplasmic reticulum stress, inflammation, and apoptosis by morin treatment. Iran J Basic Med Sci. 2025;28(3):376–84. doi:10.22038/ijbms.2024.81490.17634 PubMed PMID: 39906618; PubMed Central PMCID: PMC11790200.
39. Verhoeven F, Prati C, Chouk M, Demougeot C, Wendling D. Methotrexate and cardiovascular risk in rheumatic diseases: A comprehensive review. Expert Rev Clin Pharmacol. 2021;14(9):1105–12.
40. Tavazzi B, Lazzarino G, Di Pierro D, Giardina B. Malondialdehyde production and ascorbate decrease are associated to the reperfusion of the isolated postischemic rat heart. Free Radic Biol Med. 1992;13(1):75–8.
41. Welbat JU, Naewla S, Pannangrong W, Sirichoat A, Aranarochana A, Wigmore P. Neuroprotective effects of hesperidin against methotrexate-induced changes in neurogenesis and oxidative stress in the adult rat. Biochem Pharmacol. 2020 Aug;178:114083. doi:10.1016/j.bcp.2020.114083
42. Felemban SG, Aldubayan MA, Alhowail AH, Almami IS. Vitamin B17 Ameliorates Methotrexate-Induced Reproductive Toxicity, Oxidative Stress, and Testicular Injury in Male Rats. Mendoza-Núñez VM, editor. Oxid Med Cell Longev. 2020 Oct 27;2020:1–11. doi:10.1155/2020/4372719
43. Abdulwahhab RQ, Ali Alabdali SM. Study of the protective effects of cyanocobalamin on methotrexate induced nephrotoxicity in rats. F1000Research. 2022 Oct 31;11:1012. doi:10.12688/f1000research.124081.2
44. Öz MA, Şimşek H, Özdemir S, Küçükler S, Kandemir Ö, Mutlu H, Yazıcı R, Kandemir FM. Nephroprotective Effects of Vanillic Acid on Bortezomib‐Induced Renal Injury: Role of Oxidative Stress, Mitochondrial Dysfunction, Apoptosis and miR‐204/Nephrin/Podocin/CD2AP Axis. Basic Clin Pharmacol Toxicol. 2026 Apr;138(4):e70223. doi:10.1111/bcpt.70223
45. Çelik H, Kucukler S, Çomaklı S, Caglayan C, Özdemir S, Yardım A, Karaman M, Kandemir FM. Neuroprotective effect of chrysin on isoniazid-induced neurotoxicity via suppression of oxidative stress, inflammation and apoptosis in rats. Neurotoxicology. 2020;81:197–208.
46. Aygörmez S, Küçükler S, Çomaklı S, Şimşek H, Kandemir Ö, Kandemir FM, Maraşlı Ş. Protective Role of Polydatin Against Vancomycin‐Induced Lung Toxicity via Oxidative Stress, Inflammation, Endoplasmic Reticulum Stress, Apoptosis, and Ferroptosis Pathways. J Appl Toxicol. 2026 Apr 10;jat.70203. doi:10.1002/jat.70203
47. Akay M, Küçükler S, Özdemir S, Çomaklı S, Kandemir Ö, Şimşek H, Mutlu H, Kandemir FM. Rutin attenuates colistin-induced nephrotoxicity through modulation of inflammation, oxidative stress, and miRNA-Associated pathways: An experimental study. Food Biosci. 2026 Apr;78:108527. doi:10.1016/j.fbio.2026.108527
48. Wang SC, Huang YC, Hsiao CC, Sheen JM, Huang LT, Lo WS, Hsieh HY, Chen YC. Melatonin protects against methotrexate hepatotoxicity in young rats: Impact of PI3K/Akt/mTOR signaling. J Biochem Mol Toxicol. 2023;37(5):e23323.
49. Alqarni AM, Zeidler MP. How does methotrexate work? Biochem Soc Trans. 2020;48(2):559–67.
50. Öz MA, Şimşek H, Özdemir S, Kandemir Ö, Küçükler S, Mutlu H, Yazıcı R, Kandemir FM. Sinapic acid attenuates colistin-driven nephrotoxicity by targeting the miRNA-21/SIRT1/NF-κB pathway and facilitating tubular repair and inflammation in a rat model. Toxicol Appl Pharmacol. 2026 Feb;507:117708. doi:10.1016/j.taap.2026.117708
51. Şener G, Ekşioǧlu-Demiralp E, Çetiner M, Ercan F, Şirvanci S, Gedik N, Yeǧen BÇ. L-Carnitine ameliorates methotrexate-induced oxidative organ injury and inhibits leukocyte death. Cell Biol Toxicol. 2006 Jan;22(1):47–60. doi:10.1007/S10565-006-0025-0 PubMed PMID: 16463019.
52. Çakir T, Özkan E, Dulundu E, Topaloǧlu Ü, Şehirli AÖ, Ercan F, Şener E, Şener G. Caffeic acid phenethyl ester (CAPE) prevents methotrexate-induced hepatorenal oxidative injury in rats. J Pharm Pharmacol. 2011 Dec;63(12):1566–71. doi:10.1111/J.2042-7158.2011.01359.X PubMed PMID: 22060287.
53. Yılmaz S, Küçükler S, Şimşek H, Aygörmez S, Kandemir FM. Naringin protects against colistin-induced sciatic nerve damage by reducing oxidative stress, apoptosis and inflammation damage. Deney Ve Klin Tıp Derg. 2024;41(1):53–9.
54. Gür C, Kandemir Ö, Kandemir FM. Evaluation of the effects of chrysin on diclofenac-induced cardiotoxicity in rats by the markers of oxidative stress, endoplasmic reticulum stress and apoptosis. kvj. 2022 Jun;15(2):151–60. doi:10.30607/kvj.1062346
55. Ali YA, Ahmed AA, Abd El‐Raouf OM, Elkhoely A, Gad AM. Polydatin combats methotrexate‐induced pulmonary fibrosis in rats: Involvement of biochemical and histopathological assessment. J Biochem Mol Toxicol. 2022;36(5):e23019.
56. Akcılar R, Akcılar A, Koçak C, Koçak FE, Bayat Z, Şimşek H, Şahin S, Savran B. Effects of Ukrain on intestinal apoptosis caused by ischemia-reperfusion injury in rats. Int J Clin Exp Med. 2015;8(12):22158.
57. Öz MA, Kandemir Ö, Özdemir S, Şimşek H, Küçükler S, Yazıcı R, Mutlu H, Kandemir FM. Chrysin attenuates tramadol-driven renal dysfunction via regulation of RNA networks, antioxidant pathways, and ketogenic metabolism. Eur J Pharmacol. 2026 May;1023:178882. doi:10.1016/j.ejphar.2026.178882
58. Al-khawalde AA marddyah A, Abukhalil MH, Jghef MM, Alfwuaires MA, Alaryani FS, Aladaileh SH, Algefare AI, Karimulla S, Alasmari F, Aldal’in HK, Alanezi AA, Althunibat OY. Punicalagin Protects against the Development of Methotrexate-Induced Hepatotoxicity in Mice via Activating Nrf2 Signaling and Decreasing Oxidative Stress, Inflammation, and Cell Death. Int J Mol Sci. 2022 Oct 15;23(20):12334. doi:10.3390/ijms232012334
59. Aydin M, Cevik A, Kandemir F, Yuksel M, Apaydin A. Evaluation of hormonal change, biochemical parameters, and histopathological status of uterus in rats exposed to 50-Hz electromagnetic field. Toxicol Ind Health. 2009 Apr;25(3):153–8. doi:10.1177/0748233709102717
60. Zhao XJ, Yu HW, Yang YZ, Wu WY, Chen TY, Jia KK, Kang LL, Jiao RQ, Kong LD. Polydatin prevents fructose-induced liver inflammation and regulate Keap1/Nrf2 pathway. Redox Biol. 2018 Sep 1;18:124–37. doi:10.1016/J.REDOX.2018.07.002 PubMed PMID: 30014902.
61. Wang HL, Gao JP, Han YL, Xu X, Wu R, Gao Y, Cui XH. Comparative studies of polydatin and resveratrol on mutual transformation and antioxidative effect in vivo. Phytomedicine. 2015 May 15;22(5):553–9. doi:10.1016/J.PHYMED.2015.03.014 PubMed PMID: 25981921.
62. Uraz S, Tahan V, Aygun C, Eren F, Unluguzel G, Yuksel M, Senturk O, Avsar E, Haklar G, Celikel C, Hulagu S, Tozun N. Role of ursodeoxycholic acid in prevention of methotrexate-induced liver toxicity. Dig Dis Sci. 2008 Apr 13;53(4):1071–7. doi:10.1007/S10620-007-9949-3/FIGURES/7 PubMed PMID: 17934844.
63. Mazur AJ, Nowak D, Mannherz HG, Malicka-Błaszkiewicz M. Methotrexate induces apoptosis in CaSki and NRK cells and influences the organization of their actin cytoskeleton. Eur J Pharmacol. 2009 Jun 24;613(1–3):24–33. doi:10.1016/J.EJPHAR.2009.04.020 PubMed PMID: 19383496.
64. Jahovic N, Çevik H, Şehirli AÖ, Yeǧen BÇ, Şener G. Melatonin prevents methotrexate-induced hepatorenal oxidative injury in rats. J Pineal Res. 2003 May 1;34(4):282–7. doi:10.1034/J.1600-079X.2003.00043.X PubMed PMID: 12662351.
65. Erfan OS, Hany |, Sonpol MA, Marwa |, El-Kader A, Hany C. Protective effect of rapamycin against acrylamide-induced hepatotoxicity: The associations between autophagy, apoptosis, and necroptosis. Anat Rec. 2021 Sep 1;304(9):1984–98. doi:10.1002/AR.24587 PubMed PMID: 33480149.
66. Gencer S, Akaras N, Şimşek H, Gür C, İleritürk M, Küçükler S, Kandemir FM. The Protective Effects of Chrysin on Acrylamide-Induced Hepatotoxicity: Insights Into Oxidative Stress, Inflammation, Apoptosis, Autophagy, and Histological Evaluation in Rats. J Biochem Mol Toxicol. 2025;39(6):e70334. doi:10.1002/jbt.70334
67. Hemeida RAM, Mohafez OM. Curcumin attenuates methotraxate-induced hepatic oxidative damage in rats. J Egypt Natl Cancer Inst. 2008 Jun 1;20(2):141–8. PubMed PMID: 20029470.
68. Araya LE, Soni IV, Hardy JA, Julien O. Deorphanizing Caspase-3 and Caspase-9 Substrates In and Out of Apoptosis with Deep Substrate Profiling. ACS Chem Biol. 2021 Nov 19;16(11):2280–96. doi:10.1021/acschembio.1c00456
69. Anson F, Thayumanavan S, Hardy JA. Exogenous Introduction of Initiator and Executioner Caspases Results in Different Apoptotic Outcomes. JACS Au. 2021 Aug 23;1(8):1240–56. doi:10.1021/jacsau.1c00261
70. Georgiou KR, King TJ, Scherer MA, Zhou H, Foster BK, Xian CJ. Attenuated Wnt/β-catenin signalling mediates methotrexate chemotherapy-induced bone loss and marrow adiposity in rats. Bone. 2012 Jun;50(6):1223–33. doi:10.1016/j.bone.2012.03.027
71. Fenderico N, Van Scherpenzeel R, Goldflam M, Proverbio D, Jordens I, Kralj T, Stryeck S, Bass T, Hermans G, Ullman C, Aastrup T, Gros P, Maurice M. Anti-LRP5/6 VHHs promote differentiation of Wnt-hypersensitive intestinal stem cells. Nat Commun. 2019. doi:10.2210/PDB6H15/PDB
72. González-Sancho J, Aguilera O, García J, Pendás-Franco N, Peña C, Cal S, Herreros A, Bonilla F, Muñoz A. The Wnt antagonist DICKKOPF-1 gene is a downstream target of β-catenin/TCF and is downregulated in human colon cancer. Oncogene. 2005 Feb 3;24:1098–103. doi:10.1038/sj.onc.1208303
73. Qin K, Yu M, Fan J, Wang H, Zhao P, Zhao G, Zeng W, Chen C, Wang Y, Wang A, Schwartz Z, Hong J, Song L, Wagstaff W, Haydon RC, Luu HH, Ho SH, Strelzow J, Reid RR, He TC, Shi LL. Canonical and noncanonical Wnt signaling: Multilayered mediators, signaling mechanisms and major signaling crosstalk. Genes Dis. 2024 Jan;11(1):103–34. doi:10.1016/j.gendis.2023.01.030
74. Duspara K, Bojanic K, Pejic JI, Kuna L, Kolaric TO, Nincevic V, Smolic R, Vcev A, Glasnovic M, Curcic IB, Smolic M. Targeting the Wnt Signaling Pathway in Liver Fibrosis for Drug Options: An Update. J Clin Transl Hepatol. 2021 Sep 13;000(000):000–000. doi:10.14218/JCTH.2021.00065
75. Rutt L, Orlicky D, McCullough R. Investigating the role of Wnt3a and Wnt5a as critical factors of hepatic stellate cell activation in acute toxicant-induced liver injury. Cell Biol Toxicol. 2024 Dec 21;41. doi:10.1007/s10565-024-09956-4