Quercetin ve folik asit: metotreksat kaynaklı yumurtalık hasarı üzerindeki koruyucu etkilerin karşılaştırmalı değerlendirmesi

Year 2026, Volume: 7 Issue: 1, 232 - 243, 31.01.2026

Halime Tozak Yıldız , Kübra Tuğçe Kalkan , Özge Göktepe , Ozge Cengiz Mat , Arzu Yay

https://doi.org/10.47482/acmr.1729326

Abstract

Giriş: Metotreksat (MTX), folik asit redüktaz inhibitörü olarak görev yapan ve çeşitli klinik durumlarda yaygın olarak kullanılan bir kemoterapötik ve immünsüpresif ajandır. Terapötik etkinliğine rağmen, MTX sitotoksik bir ilaç olup, fertiliteyi olumsuz etkileyebilecek oksidatif stres aracılı over hasarı gibi önemli yan etkilere sahiptir. Bu çalışma, quercetin ve folik asidin MTX kaynaklı over hasarına karşı koruyucu etkilerini değerlendirmeyi ve karşılaştırmayı amaçlamıştır.
Yöntemler: Kırk iki adet dişi Wistar albino sıçan, rastgele olarak altı gruba (n=7) ayrıldı: Kontrol, QRC (quercetin), FA (folik asit), MTX (metotreksat), MTX+QRC (metotreksat + quercetin) ve MTX+FA (metotreksat+folik asit). Metotreksat, 1 mg/kg dozunda intraperitoneal olarak 1., 3., 5. ve 7. günlerde uygulanmıştır. Quercetin (50 mg/kg) ve folik asit (20 mg/kg) ise 2., 4., 6. ve 8. günlerde oral olarak verilmiştir. 9. günde over dokuları histopatolojik, immünohistokimyasal ve biyokimyasal analizler için toplanmıştır. İstatistiksel analizler, tek yönlü ANOVA ve Kruskal-Wallis testleri ile Bonferroni ve Dunn post hoc düzeltmeleri kullanılarak yapılmış olup p<0,05 değeri anlamlı kabul edilmiştir.
Bulgular: Metotreksat, folikül dejenerasyonu, inflamasyon, vasküler konjesyon, hemoraji, ödem ve fibrozisi anlamlı şekilde artırmıştır (p<0,001) ve malondialdehit (MDA) ile pro-inflamatuvar sitokinler (TNF-α, IL-1β, IL-6) düzeylerini yükseltmiştir (p<0,001). Ayrıca anti-Müllerian hormon (AMH) düzeyleri, süperoksit dismutaz (SOD) ve katalaz (CAT) aktiviteleri ile tüm folikül sayıları, özellikle primordial foliküller, anlamlı şekilde azalmıştır (p<0,001). Hem quercetin hem de folik asit tedavileri bu etkileri hafifletmiştir. Quercetin, TNF-α ve IL-1β düzeylerini azaltmada (p<0,01) ve antioksidan enzim aktivitelerini geri kazandırmada (p<0,05) daha etkili olurken, folik asit AMH düzeylerini korumada (p<0,01) ve primordial ile graaf folikül sayısını artırmada (p<0,05) daha etkili olmuştur.
Sonuç: Metotreksat etkili bir terapötik ajan olmaya devam etse de, over toksisitesi uzun vadeli fertilite açısından potansiyel bir risk oluşturmaktadır. Mevcut bulgular, quercetin ve folik asidin MTX kaynaklı over hasarına karşı koruyucu etkiler gösterdiğini ve bunun farklı mekanizmalarla gerçekleştiğini göstermektedir: quercetin esas olarak oksidatif stres ve inflamasyonu azaltarak, folik asit ise folikülogenezi destekleyerek ve over rezervini koruyarak etkili olmaktadır.

Keywords

Over rezervi , üreme toksikolojisi , metotreksat , kuersetin , folik asit

Quercetin versus folic acid: a comparative evaluation of protective effects on methotrexate-induced ovarian damage

Abstract

Background: Methotrexate (MTX) is a chemotherapeutic and immunosuppressive agent that acts as a folic acid reductase inhibitor and is widely used in various clinical settings. Despite its therapeutic efficacy, MTX has been shown in some experimental models to induce oxidative stress–mediated damage in ovarian tissue. This study aimed to evaluate and compare the protective effects of quercetin and folic acid against MTX-induced ovarian injury.
Methods: Forty-two female Wistar albino rats were randomly divided into six groups (n=7): Control, QRC (quercetin), FA (folic acid), MTX (methotrexate), MTX+QRC (methotrexate+ folic acid), and MTX+FA (methotrexate+folic acid). Methotrexate was administered intraperitoneally at1mg/kg on days 1, 3, 5, and 7. quercetin(50mg/kg) and folic acid(20mg/kg) were administered orally on days 2, 4, 6, and 8. On day 9, ovarian tissues were collected for histopathological, immunohistochemical and biochemical analyses. Statistical analysis was conducted using one-way ANOVA and Kruskal-Wallis tests with Bonferroni and Dunn’s post hoc corrections (p<0.05).
Results: Methotrexate significantly increased follicular degeneration, inflammation, vascular congestion, hemorrhage, edema, and fibrosis (p<0.001), as well as malondialdehyde (MDA) and pro-inflammatory cytokines (TNF-α, IL-1β, IL-6) (p<0.001). It also significantly reduced anti-Mullerian hormone (AMH) levels, superoxide dismutase (SOD) and catalase (CAT) activities, and all follicle counts including primordial follicles (p<0.001). Both quercetin and folic acid treatments ameliorated these effects. Quercetin showed greater efficacy in reducing TNF-α and IL-1β levels (p<0.01) and restoring antioxidant enzyme activities(p<0.05), while folic acid was more effective in preserving AMH levels (p<0.01) and increasing primordial and Graafian follicle numbers (p<0.05).
Conclusion: Although methotrexate remains an effective therapeutic agent, its ovarian toxicity poses a potential risk to long-term fertility. The present findings indicate that quercetin and folic acid exert protective effects against MTX-induced ovarian damage through distinct mechanisms: quercetin primarily by attenuating oxidative stress and inflammation, and folic acid by supporting folliculogenesis and preserving ovarian reserve.

Keywords

Ovarian reserve , reproductive toxicology , methotrexate , quercetin , folic acid.

Ethical Statement

The rats were obtained and accommodated at the Experimental and Clinical Research Center of Erciyes University. The study was designed and conducted in compliance with the ARRIVE guidelines for in vivo animal research. Erciyes University Animal Experiments Local Ethics Committee approved our experimental guidelines (decision no: 22/174 date: 08.08.2022).

Supporting Institution

This research received no external funding.

References

• ACOG Practice Bulletin No. 193: Tubal Ectopic Pregnancy. Obstet Gynecol. 2018;131(3):613-615. doi:10.1097/AOG.0000000000002559.
• Orazulike NC, Konje JC. Diagnosis and management of ectopic pregnancy. Womens Health (Lond). 2013;9(4):373–385. doi:10.2217/whe.13.35.
• Stabile G, Romano F, Buonomo F, Zinicola G, Ricci G. Conservative treatment of interstitial ectopic pregnancy with the combination of mifepristone and methotrexate: our experience and review of the literature. Biomed Res Int. 2020;2020:8703496. doi:10.1155/2020/8703496.
• Tonick S, Conageski C. Ectopic pregnancy. Obstet Gynecol Clin North Am. 2022;49(3):537–549. doi:10.1016/j.ogc.2022.02.018.
• Kamel MY, Ahmed SM, Abdelzaher WY, Welson NN, Abdel Aziz AM. Role of IL-6/STAT3 pathway in mediating the protective effect of agomelatine against methotrexate-induced lung and intestinal tissue damage in rats. Immunopharmacol Immunotoxicol. 2022;44(1):35–46. doi:10.1080/08923973.2021.1999973.
• Kiremitli T, Kiremitli S, Akselim B, et al. Protective effect of coenzyme Q10 on oxidative ovarian and uterine damage induced by methotrexate in rats. Hum Exp Toxicol. 2021;40(9):1537-1544. doi:10.1177/09603271211002891.
• Carrillo Martinez EJ, Flores Hernández FY, Salazar Montes AM, Nario Chaidez HF, Hernández Ortega LD. Quercetin, a flavonoid with great pharmacological capacity. Molecules. 2024;29(3):1000. doi:10.3390/molecules29031000.
• Chen Z, Yuan Q, Xu G, et al. Effects of quercetin on proliferation and H2O2-induced apoptosis of intestinal porcine enterocyte cells. Molecules. 2018;23(8):2012. doi:10.3390/molecules23082012.
• Aldemir M, Özgün G, Önen E, Okulu EM, Kayıgil Ö. Quercetin has a protective role on histopathological findings in testicular ischaemia–reperfusion injury in rats. Andrologia. 2012;44(Suppl 1):479–483. doi:10.1111/j.1439-0272.2011.01210.x.
• Wang J, Zhu H, Wang K, Yang Z, Liu Z. Protective effect of quercetin on rat testes against cadmium toxicity by alleviating oxidative stress and autophagy. Environ Sci Pollut Res Int. 2020;27(24):25278–25286. doi:10.1007/s11356-020-09029-4.
• Zheng S, Ma M, Chen Y, Li M. Effects of quercetin on ovarian function and regulation of the ovarian PI3K/Akt/FoxO3a signalling pathway and oxidative stress in a rat model of cyclophosphamide-induced premature ovarian failure. Basic Clin Pharmacol Toxicol. 2022;130(2):240-253. doi:10.1111/bcpt.13696.
• Sijilmassi O. Folic acid deficiency and vision: a review. Graefes Arch Clin Exp Ophthalmol. 2019;257(8):1573-1580. doi:10.1007/s00417-019-04344-0.
• Greenberg JA, Bell SJ, Guan Y, Yu YH. Folic acid supplementation and pregnancy: more than just neural tube defect prevention. Rev Obstet Gynecol. 2011;4(2):52-59.
• Low PS, Kularatne SA. Folate-targeted therapeutic and imaging agents for cancer. Curr Opin Chem Biol. 2009;13(3):256-262. doi:10.1016/j.cbpa.2009.04.015.
• Xin P, Xu X, Deng C, et al. The role of JAK/STAT signaling pathway and its inhibitors in diseases. Int Immunopharmacol. 2020;80:106210. doi:10.1016/j.intimp.2020.106210.
• Pahwa R, Goyal A, Bansal P, Jialal I. Chronic inflammation. In: StatPearls [Internet]. Treasure Island (FL): StatPearls Publishing; 2023.
• Germolec DR, Shipkowski KA, Frawley RP, Evans E. Markers of inflammation. In: DeWoskin RS, editor. Immunotoxicity Testing: Methods and Protocols. Vol 1803. New York: Springer; 2018. p. 57–79.
• Pandya MJ, Panchal D, Shah H. Role of methotrexate in ectopic pregnancy. Int J Reprod Contracept Obstet Gynecol. 2021;10(1):84-88.
• Beguin C, Brichant G, De Landsheere L, et al. Use of methotrexate in the treatment of ectopic pregnancies: a retrospective single center study. Facts Views Vis Obgyn. 2020;11(4):329-335.
• Nagy ZB, Barták BK, Zsigrai S, et al. Effect of DNA fragments and nucleotide supplementation on tumor development in mice model. Cancer Res. 2020;80(16 Suppl):2761.
• Rao SS, Sharma S, Dubey A, et al. Effect of quercetin and curcumin in rats sub-acutely exposed to cadmium chloride: hematobiochemical changes, oxidative stress parameters, and histopathological changes in intestine, liver, and kidney. J Phytomedicine. 2021;10(5):399–408.
• Gibson-Corley KN, Olivier AK, Meyerholz DK. Principles for valid histopathologic scoring in research. Vet Pathol. 2013;50(6):1007-1015. doi:10.1177/0300985813485099.
• Hamed KM, Dighriri IM, Baomar AF, et al. Overview of methotrexate toxicity: a comprehensive literature review. Cureus. 2022;14(9):e29171. doi:10.7759/cureus.29171.
• Fu J, Liu Y, Wang C, et al. Persistent follicular granulosa cell senescence and apoptosis induced by methotrexate leading to oocyte dysfunction and aberrant embryo development. Clin Transl Sci. 2021;14(5):2043-2054.
• Kamel WH, Ali MF, Afifi SH. Histopathological and biochemical studies of methotrexate hepatotoxicity on albino rats. Assiut Vet Med J. 2023;69(179):60-68.
• Hussein A, Ali SM. A short-term comparison between the effects of two different concentrations of methotrexate on ovarian tissues and function in female albino rats. J Fac Med Baghdad. 2023;64(4):280–285.
• Angın AD, Kaya M, Sönmez S, et al. The effect of dehydroepiandrosterone on ovarian reserve in ovarian damage caused by methotrexate. Med Lab Technol J. 2020;6(1):1-8.
• Chen Y, Zhao Y, Miao C, et al. Quercetin alleviates cyclophosphamide-induced premature ovarian insufficiency in mice by reducing mitochondrial oxidative stress and pyroptosis in granulosa cells. J Ovarian Res. 2022;15:138. doi:10.1186/s13048-022-01080-3.
• Zhang Q, He W, Wang H, et al. Folic acid preconditioning alleviated radiation-induced ovarian dysfunction in female mice. Front Nutr. 2022;9:854655.
• Asbaghi O, Ashtary-Larky D, Bagheri R, et al. Effects of folic acid supplementation on inflammatory markers: a GRADE-assessed systematic review and dose–response meta-analysis of randomized controlled trials. Nutrients. 2021;13(7):2327. doi:10.3390/nu13072327.
• Ezhilarasan D. Hepatotoxic potentials of methotrexate: understanding the possible toxicological molecular mechanisms. Toxicology. 2021;458:152840. doi:10.1016/j.tox.2021.152840.
• Elkady M, Shalaby S, Fathi F, El Mandouh S. Effects of quercetin and rosuvastatin each alone or in combination on cyclophosphamide-induced premature ovarian failure in female albino mice. Hum Exp Toxicol. 2019;38(11):1283-1295. doi:10.1177/0960327119865588.