The Protective Effects of Receptor-Interactive Protein Kinase-3 Inhibitor - Dabrafenib on Cisplatin Induced Nephrotoxicity

Abstract

Objective: The aim of this study is to investigate the protective effects of dabrafenib, a receptor-interacting kinase-3 enzyme inhibitor, against necroptosis which has an important role in Cisplatin-induced nephrotoxicity. Methods: 32 male Sprague-Dawley rats were divided into 4 groups. A single dose of i.p. saline was given to the first group. In the second group, a single dose of cisplatin (5 mg/kg i.p.) was given. In the third group, 10mg/kg i.p injections of Dabrafenib began a day before injection of a single dose of 5 mg/kg, i.p. cisplatin for 7 days. In the forth group 10mg/kg Dabrafnib was injected i.p. for 7 days. After 7 days, in all groups rats were anesthetized and with a laparotomy left kidneys were isolated. Histopathological examinations, Western-Blot analyses (expression levels of RIP1, RIP3, and MLKL proteins), and malondialdehyde (MDA) measurements were performed on the isolated kidney tissues Results: In the biochemical evaluation of the study, it was observed that MDA levels in the kidney tissue increased significantly in the Cisplatin group compared to the control group (p<0.05). This increase was evaluated as an important indicator of oxidative stress. On the other hand, a significant decrease was detected in MDA levels in the group administered Dabrafenib + Cisplatin. This decrease was found to be substantial when compared to the Cisplatin group (p<0.05). Histopathological analysis revealed that cisplatin caused serious kidney damage, but its administration with dabrafenib significantly reduced this damage and provided a protective effect (p<0.05). In addition, the expression levels of RIP1, RIP3, and MLKL proteins, which are associated with necroptosis, were examined by Western Blot analyses. Significant increases were observed in the expression levels of these proteins in the Cisplatin group compared to the control group (p<0.05). However, a significant decrease was observed in the expression levels of these proteins in Cisplatin + Dabrafenib group (p<0.05). This decrease indicates the inhibitory effect of Dabrafenib on the necroptotic pathway. Conclusion: This study suggests that the use of Dabrafenib with Cisplatin can be considered as a potential strategic agent to reduce chemotherapy-induced nephrotoxicity. However, advanced preclinical and clinical studies are needed before clinical practice.

Keywords

• Cisplatin
• Nephrotoxicity
• Dabrafenib

Reseptör Etkileşimli Protein Kinaz-3 Inhibitörü Dabrafenib’in Sisplatin Nefrotoksisitesi Üzerine Koruyucu Etkileri1

Abstract

Amaç: Bu çalışma reseptör etkileşimli kinaz-3 enzim inhibitörü olan dabrafenibin sisplatin tarafından tetiklenen nefrotoksisitede oluşan nekroptoza karşı koruyucu etkilerini araştırmaktır. Yöntemler: 32 adet Sprague-Dawley cinsi erkek rat 4 gruba ayrıldı. 1.gruba tek doz i.p. salin verildi. 2.gruba 5 mg/kg tek doz sisplatin i.p. olarak verildi. 3.gruba tek doz sisplatin (5 mg/kg,i.p) enjeksiyonundan bir gün önce başlanarak 7 gün boyunca Dabrafenib günde 10 mg/kg,i.p olarak verildi. 4. Gruba 7 gün boyunca Dabrafenib günde 10 mg/kg, i.p olarak verildi. Tüm gruplarda 7. Günün sonunda anestezi altında laparotomi yapılarak börekler izole edildi. Alınan böbrek dokularında histopatolojik incelemeler, Western-Blot analizleri (RIP1, RIP3, ve MLKL proteinlerinin ekspresyon seviyeleri) ve malondialdehit (MDA) ölçümler yapıldı. Bulgular: Çalışmada MDA düzeyleri değerlendirildiğinde, sisplatin uygulanan grupta kontrol grubuna kıyasla böbrek dokusundaki MDA düzeylerinin anlamlı düzeyde arttığı gözlendi (p<0,05). Bu artış oksidatif stresin önemli bir göstergesi olarak değerlendirildi. Buna karşın sisplatin+ dabrafenib uygulanan grupta ise MDA düzeylerinde anlamlı bir azalma saptandı ve bu azalma sisplatin grubu ile kıyaslandığında anlamlı bulundu (p<0,05). Bu durum, dabrafenibin oksidatif hasar üzerindeki hafifletici etkisini göstermektedir. Nekroptozis ile ilişkili olan RIP1, RIP3 ve MLKL proteinlerinin ekspresyon seviyeleri Western Blot analizleri ile incelendi. Sisplatin verilen grupta, kontrol grubuna göre bu proteinlerin ekspresyon düzeylerinde anlamlı artışlar görüldü (p<0,05). Bu bulgular sisplatinin nekroptozu indüklediğini göstermektedir. Ancak sisplatin+dabrafenib grubunda, bu proteinlerin ekspresyon düzeylerinde anlamlı bir azalma görüldü (p<0,05). Bu azalma da Dabrafenibin nekroptotik yolağı inhibe edici etkisine işaret etmektedir. Sonuç: Bu çalışma sisplatin ile birlikte Dabrafenib kullanımının kemoterapiye bağlı nefrotoksisiteyi azaltmak amacıyla potansiyel bir stratejik ajan olarak değerlendirilebileceğini düşündürmektedir. Ancak bu verilerin klinik uygulamaya yansıması için ileri düzeyde preklinik ve klinik çalışmalara ihtiyaç duyulmaktadır.

Keywords

• Sisplatin
• Dabrafenib
• Nefrotoksisite.

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