EVALUATION OF THE PROTECTIVE EFFECT OF RAMELTEON IN METHOTREXATE INDUCED NEPHROTOXICITY

Abstract

Objective Methotrexate (MTX), which is used as an immunosuppressive and anticancer drug, causes serious toxic side effects in many organs, including the kidney. Activation of apoptotic pathways through oxidative stress is involved in the mechanism of MTX mediated nephrotoxicity. In our study, we investigated the protective effects of ramelteon (RML), an analogue of melatonin, whose antioxidant and antiapoptotic properties are well known, on MTX nephrotoxicity. Material and Method 32 rats were divided into 4 groups as Control, MTX, MTX+RML and RML. According to the groups, saline or RML (10 mg/kg) was administered by oral gavage for 7 days, and on the 2nd day, 20 mg of MTX or the same volume of saline was administered intraperitoneally according to the groups. At the end of the experiment, the rats were sacrificed and kidney tissues were examined histopathologically with Hematoxylin-Eosin (HE) staining and immunohistochemically (IHC) with caspase-3 and TNF-α staining. In addition, serum BUN, creatinine levels were measured, kidney Total Oxidant and Antioxidant Status (TAS, TOS) levels were studied and Oxidative Stress Index (OSI) was calculated. Results Creatinine, TOS and OSI levels in the MTX group were found to be significantly higher than in the control group. In HE staining, tissue damage was significantly higher in MTX group compared to the control group, and cas-3 and TNF-α staining levels were increased in IHC staining. These findings were found to be reversed in the MTX+RML group. Conclusion We show that RML treatment improves the findings of MTX-induced nephrotoxicity. RML may be a promising drug in MTX nephrotoxicity.

Keywords

• Melatonin analogs
• drug induced nephrotoxicity
• oxidative stress
• apoptosis

METOTREKSAT ARACILI NEFROTOKSİSİTEDE RAMELTEONUN KORUYUCU ETKİSİNİN DEĞERLENDİRİLMESİ

Abstract

Amaç İmmünsüpresif ve antikanser olarak kullanılan Metotreksat (MTX), böbrek dahil birçok organda ciddi toksik yan etkilere neden olmaktadır. Mtx aracılı nefrotoksisitenin mekanizmasında oksidatif stres üzerinden apoptotik yolakların aktive olması yer almaktadır. Çalışmamızda antioksidan ve antiapoptotik özellikleri iyi bilinen melatonin’in analoglarından Ramelteon’un (RML) MTX nefrotoksisitesindeki koruyucu etkilerini araştırdık. Gereç ve Yöntem 32 adet sıçan Kontrol, MTX, MTX+RML ve RML olmak üzere 4 gruba ayrıldı. Gruplara göre 7 gün boyunca oral gavajla salin (SF) ya da RML (10 mg/kg) uygulandı, 2. gün ise gruplara göre intraperitoneal 20 mg MTX ya da aynı hacimde salin uygulandı. Deney sonunda ratlar sakrifiye edilerek böbrek dokuları Hematoksilen-Eozin (HE) boyama ile histopatolojik olarak, caspase-3 ve TNF-α boyama ile immünohistokimyasal (İHC) olarak incelendi. Ayrıca serum BUN, kreatinin düzeyleri ölçüldü ve böbrek Total oksidan ve antioksidan durum (TAS, TOS) düzeyleri çalışılarak Oksidatif stres indeksi (OSİ) hesaplandı. Bulgular MTX grubunda kreatinin, TOS ve OSİ düzeyleri Kontrol grubuna göre anlamlı düzeyde yüksek saptandı. HE boyamada MTX grubunda Kontrol grubuna göre anlamlı düzeyde yüksek doku hasarı, İHC boyamada cas-3 ve TNF-α boyanma düzeylerinde artış saptandı. Bu bulguların MTX+RML grubunda geriye çevrildiği saptandı. Sonuç RML tedavisinin, MTX’in yol açtığı nefrotoksisiteye ilişkin bulguları iyileştirdiğini gösterdik. RML, MTX nefrotoksisitesinde umut vadeden bir ilaç olabilir.

Keywords

• Apoptoz
• İlaca bağlı nefrotoksisite
• Melatonin analogları
• Oksidatif stres

References

1. 1. Cronstein BN, Aune TM. Methotrexate and its mechanisms of action in inflammatory arthritis. Nature reviews Rheumatology. 2020;16(3):145-54.
2. 2. Gaies E, Jebabli N, Trabelsi S, Salouage I, Charfi R, Lakhal M, et al. Methotrexate side effects: review article. J Drug Metab Toxicol. 2012;3(4):1-5.
3. 3. Andankar P, Shah K, Patki V. A review of drug-induced renal injury. Journal of Pediatric Critical Care. 2018;5(2):36.
4. 4. Iqbal S, Armaghani A, Aiyer R, Kazory A. Methotrexate nephrotoxicity: Novel treatment, new approach. Journal of Oncology Pharmacy Practice. 2013;19(4):373-6.
5. 5. Erdbrügger U, De Groot K. Is methotrexate nephrotoxic? Dose- dependency, comorbidities and comedication. Zeitschrift fur Rheumatologie. 2011;70(7):549-52.
6. 6. Heidari R, Ahmadi A, Mohammadi H, Ommati MM, Azarpira N, Niknahad H. Mitochondrial dysfunction and oxidative stress are involved in the mechanism of methotrexate-induced renal injury and electrolytes imbalance. Biomedicine & Pharmacotherapy. 2018;107:834-40.
7. 7. Devrim E, Çetin R, Kılıçoğlu B, Imge Ergüder B, Avcı A, Durak İ. Methotrexate causes oxidative stress in rat kidney tissues. Renal failure. 2005;27(6):771-3.
8. 8. Osman AT, Sharkawi SM, Hassan MI, Abo-Youssef AM, Hemeida RA. Empagliflozin and neohesperidin protect against methotrexate-induced renal toxicity via suppression of oxidative stress and inflammation in male rats. Food and Chemical Toxicology. 2021;155:112406.

9. 9. Amaral FGd, Cipolla-Neto J. A brief review about melatonin, a pineal hormone. Archives of endocrinology and metabolism. 2018;62:472-9.
10. 10. Reiter RJ, Tan DX, Rosales-Corral S, Galano A, Zhou XJ, Xu B. Mitochondria: central organelles for melatonin′ s antioxidant and anti-aging actions. Molecules. 2018;23(2):509.
11. 11. Pandi-Perumal SR, Srinivasan V, Poeggeler B, Hardeland R, Cardinali DP. Drug insight: the use of melatonergic agonists for the treatment of insomnia—focus on ramelteon. Nature Clinical Practice Neurology. 2007;3(4):221-8.
12. 12. Erel O. A novel automated direct measurement method for total antioxidant capacity using a new generation, more stable ABTS radical cation. Clinical biochemistry. 2004;37(4):277-85.
13. 13. Erel O. A new automated colorimetric method for measuring total oxidant status. Clinical biochemistry. 2005;38(12):1103-11.
14. 14. Yanik M, Erel O, Kati M. The relationship between potency of oxidative stress and severity of depression. Acta europsychiatrica. 2004;16(4):200-3.
15. 15. Refaiy A, Muhammad E, ElGanainy E. Semiquantitative smoothelin expression in detection of muscle invasion in transurethral resection and cystectomy specimens in cases of urinary bladder carcinoma. African Journal of Urology. 2011;17(1).
16. 16. McNaughton L, Puttagunta L, Martinez-Cuesta MA, Kneteman N, Mayers I, Moqbel R, et al. Distribution of nitric oxide synthase in normal and cirrhotic human liver. Proceedings of the National Academy of Sciences. 2002;99(26):17161-6.
17. 17. Sinha K, Das J, Pal PB, Sil PC. Oxidative stress: the mitochondria- dependent and mitochondria-independent pathways of apoptosis. Archives of toxicology. 2013;87(7):1157-80.
18. 18. Hempel L, Misselwitz J, Fleck C, Kentouche K, Leder C, Appenroth D, et al. Influence of high‐dose methotrexate therapy (HD‐MTX) on glomerular and tubular kidney function. Medical and Pediatric Oncology: The Official Journal of SIOP—International Society of Pediatric Oncology (Societé Internationale d'Oncologie Pédiatrique. 2003;40(6):348-54.
19. 19. Elmore S. Apoptosis: a review of programmed cell death. Toxicologic pathology. 2007;35(4):495-516.
20. 20. Mahmoud AM, Hussein OE, Abd El-Twab SM, Hozayen WG. Ferulic acid protects against methotrexate nephrotoxicity via activation of Nrf2/ARE/HO-1 signaling and PPARγ, and supp- ression of NF-κB/NLRP3 inflammasome axis. Food & Function. 2019;10(8):4593-607.
21. 21. Zelová H, Hošek J. TNF-α signalling and inflammation: interactions between old acquaintances. Inflammation Research. 2013;62(7):641-51.
22. 22. Campbell MT, Dagher P, Hile KL, Zhang H, Meldrum DR, Rink RC, et al. Tumor necrosis factor-α induces intrinsic apoptotic signaling during renal obstruction through truncated bid activation. The Journal of urology. 2008;180(6):2694-700.
23. 23. Türk E, Güvenç M, Cellat M, Uyar A, Kuzu M, Ağgül AG, et al. Zingerone protects liver and kidney tissues by preventing oxidative stress, inflammation, and apoptosis in methotrexate-treated rats. Drug and Chemical Toxicology. 2020:1-12.
24. 24. Ferlazzo N, Andolina G, Cannata A, Costanzo MG, Rizzo V, Currò M, et al. Is melatonin the cornucopia of the 21st century? Antioxidants. 2020;9(11):1088.
25. 25. Oleshchuk O, Ivankiv Y, Falfushynska H, Mudra A, Lisnychuk N. Hepatoprotective effect of melatonin in toxic liver injury in rats. Medicina. 2019;55(6):304.
26. 26. Haghi-Aminjan H, Asghari MH, Farhood B, Rahimifard M, Hashemi Goradel N, Abdollahi M. The role of melatonin on chemotherapy-induced reproductive toxicity. Journal of Pharmacy and Pharmacology. 2018;70(3):291-306.
27. 27. Moradi M, Goodarzi N, Faramarzi A, Cheraghi H, Hashemian AH, Jalili C. Melatonin protects rats testes against bleomycin, etoposide, and cisplatin-induced toxicity via mitigating nitro-oxidative stress and apoptosis. Biomedicine & Pharmacotherapy. 2021;138:111481.
28. 28. Fisher SP, Davidson K, Kulla A, Sugden D. Acute sleep‐promoting action of the melatonin agonist, ramelteon, in the rat. Journal of pineal research. 2008;45(2):125-32.
29. 29. Jahovic N, Cevik H, Şehirli AÖ, Yeğen BÇ, Şener G. Melatonin prevents methotrexate‐induced hepatorenal oxidative injury in rats. Journal of pineal research. 2003;34(4):282-7.
30. 30. Abraham P, Kolli VK, Rabi S. Melatonin attenuates methotrexate‐ induced oxidative stress and renal damage in rats. Cell biochemistry and function. 2010;28(5):426-33.
31. 31. Oguz E, Kocarslan S, Tabur S, Sezen H, Yilmaz Z, Aksoy N. Effects of lycopene alone or combined with melatonin on methotrexate-induced nephrotoxicity in rats. Asian Pacific journal of cancer prevention. 2015;16(14):6061-6.