THE AMELIORATING EFFECTS OF BILBERRY ON METHOTREXATE-INDUCED INTESTINAL INJURY

Öz

Objective: Methotrexate (MTX) is a folic acid analogue chemotherapeutic used in the treatment of some malignant tumors and autoimmune diseases. In addition to its antitumoral properties, it can also exhibit toxic effects on the kidney and intestines. Bilberry (BB) fruit is a potent natural antioxidant rich in anthocyanidins. The purpose of this study was to investigate the potential ameliorating effects of BB against MTX-induced intestinal damage using biochemical and histological methods. Material and Methods: Twenty-one adult female Sprague Dawley rats were divided into three equal groups. No procedure was performed on the control group (ControlG), while the MTX group (MTXG) and MTX+BB extract group (MTX+BBG) received a single intraperitoneal dose of 30 mg/kg MTX on the first day of the experiment. MTX+BBG also received 200 mg/kg BB extract by oral gavage once daily for five days starting on the first day of the experiment. Half the intestinal tissues removed from the duodenal region at the experiment were used for biochemical evaluation, and the other half for histological examination. Results: Malondialdehyde (MDA), total oxidant status (TOS), oxidative stress index (OSI), and 8-oxo-2′-deoxyguanosine (8- OHdG) values were all higher in MTXG intestinal tissues than in ControlG. MDA, TOS, OSI, and 8-OHdG values were lower in MTX+BBG than in MTXG. In addition, caspase-3 levels in MTXG were higher than those in both ControlG and MTX+BBG, while total antioxidant status (TAS) values were lower. In terms of histology, villous hemorrhage, inflammatory cell infiltration, fusion, and degeneration in the villus epithelium were present in MTXG intestinal tissue, and the total damage score was also high. Improvement in all these parameters was present in MTX+BBG. Conclusion: Orally administered BB extract can improve MTX-induced intestinal damage through its antioxidant and anti-inflammatory effects.

Anahtar Kelimeler

• Antioxidant
• intestine
• methotrexate
• oxidative stress
• bilberry

Metotreksatın İndüklediği İntestinal Hasar Üzerine Yaban Mersininin İyileştirici Etkileri

Öz

Amaç: Metotreksat (MTX), bazı malign tümörlerin ve otoimmün hastalıkların tedavisinde kullanılan folik asit analoğu bir kemoterapötiktir. Antitümoral etkilerinin yanında karaciğer, böbrek ve bağırsaklar üzerine toksik etkiler de gösterebilir. Yaban mersini (YM), güçlü ve doğal bir antioksidan olan antosiyaninlerden zengin bir meyvedir. Bu çalışma; MTX ile indüklenen intestinal hasara karşı YM ekstraktının olası iyileştirici etkilerini biyokimyasal ve histolojik olarak araştırmayı amaçlamaktadır. Gereç ve Yöntemler: Çalışmada kullanılan 21 adet Spraque Dawley ırkı erişkin dişi sıçan, 3 eşit gruba ayrıldı. Kontrol grubu (KontrolG)’na hiçbir işlem uygulanmadı. MTX grubu (MTXG)’na ve MTX+YM ekstraktı grubu (MTX+YMG)’na deneyin ilk günü tek doz 30 mg/kg MTX intraperitoneal olarak uygulandı. MTX+YMG’na, ilave olarak deneyin ilk gününden itibaren 5 gün, günde bir kez oral gavaj ile 200 mg/kg YM ektraktı uygulandı. Deney sonunda duodenal bölgeden alınan intestinal dokuların yarısı biyokimyasal, diğer yarısı histolojik olarak değerlendirildi. Bulgular: MTXG bağırsak dokularında; malondialdehid (MDA), total oksidan kapasite (TOS), oksidatif stres indeksi (OSI) ve 8-oxo-2′-deoxyguanosine (8-OHdG) değerleri KontrolG’na göre yüksekti. MTX+YMG’nun MDA, TOS, OSI ve 8-OHdG değerleri ise MTXG’na göre azalmıştı. İlave olarak MTXG’nun Kaspaz 3 seviyesi hem ControlG’na hem MTX+YMG’na göre yüksek; total antioksidan durum (TAS) seviyeleri ise düşüktü. Histolojik olarak MTXG bağırsak dokusunun villuslarında hemoraji, inflamatuvar hücre infiltrasyonu, füzyon ve villus epitelinde dejenerasyon vardı ve total hasar skoru da yüksekti. MTX+YMG’nda ise tüm bu patolojilerde hafifleme mevcuttu. Sonuç: Oral olarak uygulanan YM ekstraktı; antioksidan ve antienflamatuvar etkileri ile MTX’in indüklediği bağırsak hasarını iyileştirebilir.

Anahtar Kelimeler

• Antioksidan
• bağırsak
• metotreksat
• oksidatif stress
• yaban mersini

Kaynakça

1. Akbarali HI, Muchhala KH, Jessup DK, Cheatham S. Chemotherapy induced gastrointestinal toxicities. Adv Cancer Res. 2022;155:131-66.
2. Marin GE, Neag MA, Burlacu CC, Buzoianu AD. The protective effects of nutraceutical components in methotrexate-induced toxicity models-An overview. Microorganisms. 2022;10(10):2053.
3. Hanoodi M, Mittal M. Methotrexate. In: Aboubakr S, Abu-Ghosh A, Sedeh PA, Aebi TC, Aeddula NR, Agadi S et al, eds. StatPearls, Treasure Island (FL): StatPearls Publishing, 2023.
4. Çataklı D, Sevük M, Coşan S, İmeci O, Keskin E, Sevinç Y, et al. Ramelteon protects intestinal tissue against injury caused by methotrexate via showing anti-apoptotic, antiinflammatory and antioxidant properties. Bezmiâlem Science. 2022;10(6):770-6.
5. Manna K, Khan ZS, Saha M, Mishra S, Gaikwad N, Bhakta JN, et al. Manjari medika grape seed extract protects methotrexate-induced hepatic ınflammation: Involvement of NF-kB/NLRP3 and Nrf2/HO-1 signaling system. J Inflamm Res. 2023;16:467-92.
6. Kanpalta F, Ozbeyli D, Sen A, Cevik O, Sener G, Ercan F. Bitter mellon (Momordica charantia) fruit extract ameliorates methotrexate-induced reproductive toxicity in male rats. Marmara Med J. 2021;34:222-8.
7. Yulug E, Turedi S, Alver A, Turedi S, Kahraman C. Effects of resveratrol on methotrexate-induced intestinal injury. Bratisl Lek Listy. 2015;116(11):676-80.
8. Ozcicek F, Kara AV, Akbas EM, Kurt N, Yazici GN, Cankaya M, et al. Effects of anakinra on the small intestine mucositis induced by methotrexate in rats. Exp Anim. 2020;69(2):144-52.

9. Alakaya M, Çıtak EÇ, Taşkınlar S, Yılmaz SN, Özçimen AA, Çimen MYB. Effects of glutamine and β-hydroxy β-methyl butyrate on methotrexate induced intestinal mucositis. Cukurova Medical Journal. 2020;45(1):266-71.
10. Aldhahrani A. Protective effects of guarana (Paullinia cupana) against methotrexate-induced intestinal damage in mice. Food Sci Nutr. 2021;9(7):3397-40.
11. Yamamoto T, Machida T, Tanno C, Hasebe S, Tamura M, Kobayashi N, et al. Low-dose nafamostat mesilate ameliorates tissue injury and inhibits 5- hydroxytryptamine synthesis in the rat intestine after methotrexate administration. J Pharmacol Sci. 2023;152(2):90-102.
12. Li P, Inoue Y, Miyamoto D, Adachi T, Okada S, Adachi T, et al. Therapeutic effect and mechanism of Daikenchuto in a model of methotrexate-induced acute small intestinal mucositis. PLoS One. 2023;18(3):e0283626.
13. Chu W, Cheung SCM, Lau RAW, Benzie IFF. Chapter 4: Bilberry (Vaccinium myrtillus L.). In: Benzie IFF, Wachtel-Galor S, eds. Herbal Medicine: Biomolecular and Clinical Aspects. 2nd edition. Boca Raton (FL): CRC Press/Taylor & Francis; 2011.
14. Vaneková Z, Rollinger JM. Bilberries: Curative and miraculous. A Review on bioactive constituents and clinical research. Front Pharmacol. 2022;13:909914.
15. Sharma A, Lee HJ. Anti-Inflammatory Activity of Bilberry (Vaccinium myrtillus L.). Curr Issues Mol Biol. 2022;44(10):4570-83.
16. Özdemir D, Özgür A, Kalkan Y, Terzi S, Tümkaya L, Yılmaz A, et al. The protective effects of whortleberry extract against cisplatin-induced ototoxicity in rats. Braz J Otorhinolaryngol. 2019;85(1):55-62.
17. Ergene Ş, Hemşinli D, Karakişi SO, Mercantepe T, Tumkaya L, Yilmaz A. The role of vaccinium myrtillus in the prevention of renal injury in an experimental model of ruptured abdominal aortic aneurysm. Braz J Cardiovasc Surg. 2020;35(4):490- 7.
18. Demir S, Turan I, Demir F, Ayazoglu Demir E, Aliyazicioglu Y. Cytotoxic effect of laurocerasus officinalis extract on human cancer cell lines. Marmara Pharm J. 2017;21(1):121-6.
19. Tusat M, Mentese A, Demir S, Alver A, Imamoglu M. Medical ozone therapy reduces oxidative stress and testicular damage in an experimental model of testicular torsion in rats. Int Braz J Urol. 2017;43(6):1160-6.
20. Sougiannis AT, VanderVeen BN, Davis JM, Fan D, Murphy EA. Understanding chemotherapy-induced intestinal mucositis and strategies to improve gut resilience. Am J Physiol Gastrointest Liver Physiol. 2021;320(5):712-9.
21. Kaymakcı A, Güven Ş, Zemheri IE, Mirapoğlu SL, Arpacık M, Şahin C. The role of sildenafil and Bacillus clausii for the prevention of methotrexate induced mucositis: A preclinical study. OTJHS. 2021;6(3):426-32.
22. Koppelmann T, Pollak Y, Ben-Shahar Y, Gorelik G, Sukhotnik I. The Mechanisms of the anti- inflammatory and anti-apoptotic effects of omega-3 polyunsaturated fatty acids during methotrexate- induced intestinal damage in cell line and in a rat model. Nutrients. 2021;13(3):888.
23. Pizzino G, Irrera N, Cucinotta M, Pallio G, Mannino F, Arcoraci V, et al. Oxidative stress: Harms and benefits for human health. Oxid Med Cell Longev. 2017;2017:8416763.
24. Elmore S. Apoptosis: A review of programmed cell death. Toxicol Pathol. 2007;35(4):495-516.
25. Kim YJ, Kim EH, Hahm KB. Oxidative stress in inflammation-based gastrointestinal tract diseases: Challenges and opportunities. J Gastroenterol Hepatol. 2012;27(6):1004-10.
26. Ozlem K, Birkan Y, Mustafa K, Emin K. Protective effect of Vaccinium myrtillus on ischemia- reperfusion injury in rat ovary. Taiwan J Obstet Gynecol. 2018;57(6):836-41.
27. Osada H, Okamoto T, Kawashima H, Toda E, Miyake S, Nagai N. Neuroprotective effect of bilberry extract in a murine model of photo-stressed retina. PLoS One. 2017;12(6)e0178627.
28. Jakesevic M, Xu J, Aaby K, Jeppsson B, Ahrné S, Molin G. Effects of bilberry (Vaccinium myrtillus) in combination with lactic acid bacteria on intestinal oxidative stress induced by ischemia-reperfusion in mouse. J Agric Food Chem. 2013;61(14):3468-78.
29. Widén C, Coleman M, Critén S, Karlgren- Andersson P, Renvert S, Persson GR. Consumption of bilberries controls gingival inflammation. Int J Mol Sci. 2015;16(5):10665-73.