Effects of rosmarinic acid and quercetin on methotrexate-induced liver and small intestine damage in rats

Yıl 2024, Cilt: 49 Sayı: 3, 547 - 560, 30.09.2024

Derya Öztürk Okatan , İlyas Ercan Okatan Abdulkadir Kutlu , Elif Şahin , Neslihan Sağlam , Ahmet Alver , Engin Yenilmez

https://doi.org/10.17826/cumj.1430648

Öz

Purpose: Rosmarinic acid (RA) is a natural phenolic compound with antioxidant and anti-inflammatory effects. Quercetin (QCT) is a powerful antioxidant that prevents oxidative damage and cell death by clearing oxygen radicals. It also has anti-inflammatory effects. In this study, it was aimed to compare the effects of RA and QCT against liver and small bowel damage that may occur due to methotrexate (MTX) use.
Materials and Methods: The study was conducted on a model of MTX-induced liver and small intestine damage in 40 Spraque Dawley male rats. RA and QCT were administered separately and in combination prophylactically (MTX+QCT group, MTX+RA group, MTX+QCT+RA group respectively). At the end of the study, liver and small intestine tissue were removed. Histopathological evaluations were performed using scoring. Malondialdehyde level, superoxide dismutase (SOD) and glutathione peroxidase (GPX) activities were examined in the tissues.
Results: In the liver tissue, pathological findings of all prophylaxis groups decreased considerably. When compared to the control group, MDA level increased significantly in the MTX, MTX+RA and MTX+RA+QCT groups The SOD and GPX activities of the MTX group decreased significantly when compared to the control group. It was found that GPX activity increased in the MTX+QCT group and SOD activity increased in the MTX+QCT+RA group when compared to the MTX group. In addition, SOD activity was significantly increased in the MTX+QCT+RA group when compared to the MTX+RA and MTX+QCT groups. In the small intestine tissue, pathological findings decreased significantly in the MTX+QCT group. Pathological findings decreased slightly in MTX+RA, MTX+QCT+RA groups. MDA levels were significantly higher in the MTX and MTX+RA+QCT groups when compared to the control group. The SOD and GPX activities of the MTX group decreased significantly compared to the control group. GPX activity decreased significantly in the MTX+QCT and MTX+RA groups when compared to the control group. SOD activity increased significantly in MTX+RA+QCT group when compared to MTX group, GPX activity increased significantly in MTX+RA+QCT group compared to MTX+QCT group.
Conclusion: RA and QCT may be effective in preventing liver damage caused by MTX. It was concluded that QCT may be more effective than RA in preventing small bowel injury caused by MTX.

Anahtar Kelimeler

Rosmarinic acid, quercetin, methotrexate, liver, small intestine tissue

Etik Beyan

Ethical approval was obtained for this study from Karadeniz Technical University Animal Experiments Local Ethics Committee with Protocol no: 2022/17.

Destekleyen Kurum

This study received no financial support

Teşekkür

We presented this study as an oral presentation at the 1th International Congress of Histology and Embryology (NICHE 2022), 26-28 May 2022, Türkiye.

Rosmarinik asit ve kuersetinin sıçanlarda metotreksat kaynaklı karaciğer ve ince bağırsak hasarı üzerine etkileri

Öz

Amaç: Rosmarinik asit (RA), antioksidan ve antiinflamatuar etkileri olan doğal bir fenolik bileşiktir. Kuersetin (QCT), oksijen radikallerini temizleyerek oksidatif hasarı ve hücre ölümünü önleyen güçlü bir antioksidandır. Aynı zamanda anti-inflamatuar etkileri de vardır. Bu çalışmada metotreksat (MTX) kullanımına bağlı oluşabilecek karaciğer ve ince bağırsak hasarına karşı RA ve QCT'nin etkilerinin karşılaştırılması amaçlandı.
Gereç ve Yöntem: Çalışma, 40 Spraque Dawley erkek sıçanda MTX'in neden olduğu karaciğer ve ince bağırsak hasarı modeli üzerinde gerçekleştirildi. RA ve QCT profilaktik olarak ayrı ayrı ve kombinasyon halinde uygulandı (sırasıyla MTX+QCT grubu, MTX+RA grubu, MTX+QCT+RA grubu). Çalışmanın sonunda karaciğer ve ince bağırsak dokusu çıkarıldı. Histopatolojik değerlendirmeler skorlama kullanılarak yapıldı. Dokularda malondialdehit düzeyi, süperoksit dismutaz (SOD) ve glutatyon peroksidaz (GPX) aktiviteleri incelendi.
Bulgular: Karaciğer dokusunda tüm profilaksi gruplarının patolojik bulgularında belirgin azalma görüldü. MTX, MTX+RA ve MTX+RA+QCT gruplarında MDA düzeyi kontrol grubuna göre arttı. MTX grubunun SOD ve GPX aktiviteleri kontrol grubuna göre azaldı. MTX grubuna göre MTX+QCT grubunda GPX aktivitesinin, MTX+QCT+RA grubunda ise SOD aktivitesinin arttığı tespit edildi. MTX+QCT+RA grubunda SOD aktivitesi, MTX+RA ve MTX+QCT gruplarına göre arttı. İnce bağırsak dokusunda patolojik bulgular MTX+QCT grubunda azaldı. MTX+RA, MTX+QCT+RA gruplarında patolojik bulgular bir miktar azaldı. MTX ve MTX+RA+QCT gruplarında MDA düzeyleri kontrol grubuna göre yüksekti. MTX grubunun SOD ve GPX aktivitelerinde kontrol grubuna göre azalma görüldü. GPX aktivitesi MTX+QCT ve MTX+RA gruplarında kontrol grubuna göre azaldı. MTX+QCT+RA grubunda MTX grubuna göre SOD aktivitesinin arttığı tespit edildi. GPX aktivitesi MTX+RA+QCT grubunda MTX+QCT grubuna göre arttı.
Sonuç: MTX'in neden olduğu karaciğer hasarını önlemede RA ve QCT'nin etkili olabileceği sonucuna varıldı. MTX'in neden olduğu ince bağırsak hasarını önlemede QCT'nin RA'dan daha etkili olabileceği sonucuna varıldı.

Anahtar Kelimeler

Rosmarinik asit, kuersetin, metotreksat, karaciğer, ince bağırsak

Kaynakça

• El-Sheikh AA, Morsy MA, Al-Taher AY. Multi-drug resistance protein (Mrp) 3 may be involved in resveratrol protection against methotrexate-induced testicular damage. Life Sci. 2014;119:40-6.
• Wang Y, Zhao TT, Zhao HY & Wang H. Melatonin protects methotrexate-induced testicular injury in rats. Eur Rev Med Pharmacol Sci. 2018;22:7517-25.
• Karri S, Vanithakumari G. Effect of methotrexate and leucovorin on female reproductive tract of albino rats. Cell Biochem Funct. 2011;29:1–21.
• Gökçe A, Oktar S, Koc A & Yonden Z. Protective effects of thymoquinone against methotrexate-induced testicular injury. Hum Exp Toxicol. 2011;30:897-903.
• Widemann BC, Adamson PC. Understanding and managing methotrexate nephrotoxicity. Oncologist. 2006;11:694-703.
• Yuluğ E, Türedi S, Alver A, Türedi S, & Kahraman C. Effects of resveratrol on methotrexate-induced testicular damage in rats. ScientificWorldJournal. 2013;2013:489659.
• Armagan A, Uzar E, Uz E, Yilmaz HR, Kutluhan S, Koyuncuoglu HR et al. Caffeic acid phenethyl ester modulates methotrexate-induced oxidative stress in testes of rat. Hum Exp Toxicol. 2008;7:547-52.
• Uzar E, Koyuncuoglu HR, Uz E, Yilmaz HR, Kutluhan S, Kilbas S et al. The activities of antioxidant enzymes and the level of malondialdehyde in cerebellum of rats subjected to methotrexate: protective effect of caffeic acid phenethyl ester. Mol Cell Biochem. 2006;291:63–8.
• Ciftci O, Cetin A, Aydin M, Kaya K, Oguz, F. Fish oil, contained in eicosapentaenoic acid and docosahexaenoic acid, attenuates testicular and spermatological damage induced by cisplatin in rats. Andrologia. 2014;46:1161-8.
• Gamaro GD, Suyenaga E, Borsoi M, Lermen J, Pereira P, Ardenghi P. Effect of rosmarinic and caffeic acids on inflammatory and nociception process in rats. ISRN Pharmacol. 2011;2011:451682.
• Malo C, Gil L, Gonzalez N, Martínez F, Cano R, de Blas I et al. Anti-oxidant supplementation improves boar sperm characteristics and fertility after cryopreservation: comparison between cysteine and rosemary (Rosmarinus officinalis). Cryobiology. 2010;61:142-7.
• Koşar M, Göger F, Can Başer KH. In vitro antioxidant properties and phenolic composition of Salvia virgata Jacq. from Turkey. J Agricult Food Chem. 2008;56:2369-74.
• Dodda D, Chhajed R, Mishra J, Padhy M. Targeting oxidative stress attenuates trinitrobenzene sulphonic acid induced inflammatory bowel disease like symptoms in rats: role of quercetin. Indian J Pharmacol. 2014;46:286.
• Aldemir M, Özgün G, Önen E, Okulu E, Kayıgil, Ö. Quercetin has a protective role on histopathological findings on testicular ischaemia–reperfusion injury in rats. Andrologia. 2012;44:479-83.
• Ranawat P, Kaushik G, Saikia UN, Pathak CM, Khanduja KL. Quercetin impairs the reproductive potential of male mice. Andrologia. 2013;45:56-65.
• Liu X, Zhang Y, Liu L, Pan Y, Hu Y, Yang P et al. Protective and therapeutic effects of nanoliposomal quercetin on acute liver injury in rats. BMC Pharmacol Toxicol. 2020;21:11.
• Bancroft JD, Cook HC, Stirling RW. Manual of Histological Techniques and Their Diagnostic Applications. London, Churchill Livingstone, 1994.
• Akman AU, Erisgin Z, Turedi S, Tekelioglu Y. Methotrexate-induced hepatotoxicity in rats and the therapeutic properties of vitamin E: a histopathologic and flowcytometric research. Clin Exp Hepatol. 2023;9:359-67.
• Kerimoğlu G, Bilgin G, Kerimoğlu S, Türkmen Alemdar N, Demir S, Aliyazıcıoğlu Y. The amelıoratıng effects of bılberry on methotrexate-ınduced ıntestınal ınjury. Kırıkkale Uni Med J. 2023;25:455-62.
• Sancak EB, Akbas A, Silan C, Cakir DU, Turkon H, Ozkanli SS. Protective effect of syringic acid on kidney ischemia-reperfusion injury. Ren Fail. 2016;38:629-35.
• Mihara M, Uchiyama M. Determination of malondialdehyde precursor in tissues by thiobarbituric acid test. Anal Biochem. 1978;86:271.
• Sun Y, Oberley LW, Li Y. A simple method for clinical assay of superoxide dismutase. Clin Chem. 1988;34:497.
• Bradford MM. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976;72:248-54.
• Samdanci ET, Huz M, Ozhan O, Tanbek K, Pamukcu E, Akatli AN et al. Cytoprotective effects of molsidomine against methotrexate-induced hepatotoxicity: an experimental rat study. Drug Des Dev Ther. 2018;13:13-21.
• Ewees MG, Abdelghany TM, Abdel-Aziz AH, Abdelbakky MS. Enoxaparin prevents fibrin accumulation in liver tissues and attenuates methotrexate-induced liver injury in rats. Naunyn Schmiedebergs Arch Pharmacol. 2019;392:623-31.
• Mohamed DI, Khairy E, Tawfek SS, Habib EK, Fetouh M.A. Coenzyme Q10 attenuates lung and liver fibrosis via modulation of autophagy in methotrexate treated rat. Biomed Pharmacother. 2019;109:892-901.
• Arslan A, Ozcicek F, Keskin Cimen F, Altuner D, Yarali O, Kurt N et al. Protective effect of resveratrol against methotrexate-induced oxidative stress in the small intestinal tissues of rats. Int J Clin Exp Med. 2015;8:10491-500.
• Southcott E, Tooley KL, Howarth GS, Davidson GP, Butler RN. Yoghurts containing probiotics reduce disruption of the small intestinal barrier in methotrexate-treated rats. Dig Dis Sci. 2008;53:1837-41.
• Pınar N, Çakırca G, Özgür T, Kaplan M. The protective effects of alpha lipoic acid on methotrexate induced testis injury in rats. Biomedicine & Pharmacotherapy. 2018;97:1486-92.
• Ma JQ, Li Z, Xie WR, Liu CM, Liu SS. Quercetin protects mouse liver against CCl₄-induced inflammation by the TLR2/4 and MAPK/NF-κB pathway. Int Immunopharmacol. 2015;28:531-9.
• Ince E. The protective effect of quercetin in the alcohol-induced liver and lymphoid tissue injuries in newborns. Mol Biol Rep. 2020;47:451-59.
• Hasanein P, Seifi R. Beneficial effects of rosmarinic acid against alcohol-induced hepatotoxicity in rats. Can J Physiol Pharmacol. 2018;96:32-7.
• Pingili RB, Challa SR, Pawar AK, Toleti V, Kodali T, Koppula S. A systematic review on hepatoprotective activity of quercetin against various drugs and toxic agents: Evidence from preclinical studies. Phytother Res. 2020;34:5-32.
• Fang P, Dou B, Liang J, Hou W, Ma C, Zhang Q. Quercetin reduces oxidative stress and apoptosis by inhibiting HMGB1 and its translocation, thereby alleviating liver injury in ACLF rats. Evid Based Complement Alternat Med. 2021;2021:2898995.
• Aydın B. Quercetin Prevents Methotrexate-induced Hepatotoxicity without Interfering Methotrexate Metabolizing Enzymes in Liver of Mice. Journal of Applied Biological Sciences. 2011;5:75-80.
• Jafaripour L, Naserzadeh R, Alizamani E, Javad Mashhadi SM, Moghadam ER et al. Effects of rosmarinic acid on methotrexate-induced nephrotoxicity and hepatotoxicity in wistar rats. Indian J Nephrol. 2021;31:218-24.
• Tóth Š, Jonecová Z, Čurgali K, Maretta M, Šoltés J, Švaňa M et al. Quercetin attenuates the ischemia reperfusion induced COX-2 and MPO expression in the small intestine mucosa. Biomed Pharmacother. 2017;95:346-54.
• Zhao P, Hu Z, Ma W, Zang L, Tian Z, Hou Q. Quercetin alleviates hyperthyroidism-induced liver damage via Nrf2 Signaling pathway. Biofactors. 2020;46:608-19.
• Gheshlaghi-Ghadim A, Mohammadi V, Zadeh-Hashem E. Protective effects of quercetin on clothianidin-induced liver damage in the rat model. Evid Based Complement Alternat Med. 2022;2022:9399695.
• Devrim E, Ergüder İB, Kılıçoğlu B, Yaykaşlı E, Çetin R, Durak I. Effects of electromagnetic radiation use on oxidant/antioxidant status and DNA turn-over enzyme activities in erythrocytes and heart, kidney, liver, and ovary tissues from rats: possible protective role of vitamin C. Toxicol Mech Methods. 2008;18:679–83.
• Oğuz A, Böyük A, Ekinci A, Alabalik U, Türkoğlu A, Tuncer MC et al. Investigation of antioxidant effects of rosmarinic acid on liver, lung and kidney in rats: a biochemical and histopathological study. Folia Morphol (Warsz). 2020;79:288-95.