EVALUATION OF THE PROTECTIVE EFFECT OF RAMELTEON IN METHOTREXATE INDUCED NEPHROTOXICITY

Öz

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.

Anahtar Kelimeler

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

METOTREKSAT ARACILI NEFROTOKSİSİTEDE RAMELTEONUN KORUYUCU ETKİSİNİN DEĞERLENDİRİLMESİ

Öz

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.

Anahtar Kelimeler

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

Kaynakça

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