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

Year 2025, Volume: 52 Issue: 3, 507 - 515, 16.09.2025

Mehmet Küçüköner , Zeynep Erdoğmuş Özgen , Fesih Aktar , Ulas Alabalık , Meryem Şeyda Kaya , Levent Erdinç İlker Kelle Meral Erdinç

https://doi.org/10.5798/dicletip.1785013

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

Ethical Statement

Ethical approval for the study was obtained from the Institutional Animal Ethics Committee, decision number 1, dated January 27, 2022. The Dicle University Scientific Research Projects Unit supported this project with the grant number TIP.22.012.

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.

References

• 1.Tang C, Livingston MJ, Safirstein R, Dong Z.Cisplatin nephrotoxicity: new insights andtherapeutic implications. Nat Rev Nephrol. 2023Jan;19(1):53–72.
• 2.Manohar S, Leung N. Cisplatin nephrotoxicity: areview of the literature. J Nephrol. 2018Feb;31(1):15–25.
• 3.Zhu S, Pabla N, Tang C, He L, Dong Z. DNA damageresponse in cisplatin-induced nephrotoxicity. ArchToxicol. 2015 Dec;89(12):2197–205.
• 4.Alassaf N, Attia H. Autophagy and necroptosis incisplatin-induced acute kidney injury: Recentadvances regarding their role and therapeuticpotential. Front Pharmacol. 2023 Jan30;14:1103062.
• 5.Linkermann A. Nonapoptotic cell death in acutekidney injury and transplantation. Kidney Int. 2016Jan;89(1):46–57.
• 6.Shan B, Pan H, Najafov A, Yuan J. Necroptosis indevelopment and diseases. Genes Dev. 2018 Mar1;32(5–6):327–40.
• 7.Dhuriya YK, Sharma D. Necroptosis: a regulatedinflammatory mode of cell death. Journal ofNeuroinflammation. 2018 Jul 6;15(1):199.
• 8.Vandenabeele P, Galluzzi L, Vanden Berghe T,Kroemer G. Molecular mechanisms of necroptosis:an ordered cellular explosion. Nat Rev Mol Cell Biol.2010 Oct;11(10):700–14.
• 9.Zhou W, Yuan J. Necroptosis in health anddiseases. Semin Cell Dev Biol. 2014 Nov;35:14–23.
• 10.Li JX, Feng JM, Wang Y, et al. The B-Raf(V600E)inhibitor dabrafenib selectively inhibits RIP3 andalleviates acetaminophen-induced liver injury. CellDeath Dis. 2014 Jun 5;5(6):e1278.
• 11.Pushpan CK, Kresock DF, Ingersoll MA, et al.Repurposing AZD5438 and Dabrafenib for Cisplatin-Induced AKI. J Am Soc Nephrol. 2024 Jan 1;35(1):22–40.
• 12.Ingersoll MA, Lutze RD, Pushpan CK, et al.Dabrafenib protects from cisplatin-induced hearingloss in a clinically relevant mouse model. JCI Insight[Internet]. 2023 Dec 22 [cited 2025 Jun 16];8(24).Available from:https://insight.jci.org/articles/view/171140
• 13.Jennette JC, Olson JL, Silva FG, et al.ResearchGate. [cited 2025 Jun 24]. Scoring systemfor renal histopathology. Available from:https://www.researchgate.net/figure/Scoring-system-for-renal-histopathology_tbl1_342423429
• 14.Yao X, Panichpisal K, Kurtzman N, Nugent K.Cisplatin nephrotoxicity: a review. Am J Med Sci.2007 Aug;334(2):115–24.
• 15.Kolbrink B, von Samson-Himmelstjerna FA,Murphy JM, Krautwald S. Role of necroptosis inkidney health and disease. Nat Rev Nephrol. 2023May;19(5):300–14.
• 16.Choi ME, Price DR, Ryter SW, Choi AMK.Necroptosis: a crucial pathogenic mediator ofhuman disease. JCI Insight. 2019 Aug8;4(15):e128834, 128834.
• 17.Xu Y, Ma H, Shao J, et al. A Role for TubularNecroptosis in Cisplatin-Induced AKI. J Am SocNephrol. 2015 Nov;26(11):2647–58.
• 18.Wang JN, Liu MM, Wang F, et al. RIPK1 inhibitorCpd-71 attenuates renal dysfunction in cisplatin-treated mice via attenuating necroptosis,inflammation and oxidative stress. Clin Sci (Lond).2019 Jul 31;133(14):1609–27.
• 19.Shi Y, Huang C, Zhao Y, et al. RIPK3 blockadeattenuates tubulointerstitial fibrosis in a mousemodel of diabetic nephropathy. Sci Rep. 2020 Jun26;10(1):10458.
• 20.Lee JE, Kim JY, Leem J. Efficacy of Trametinib inAlleviating Cisplatin-Induced Acute Kidney Injury:Inhibition of Inflammation, Oxidative Stress, andTubular Cell Death in a Mouse Model. Molecules.2024 Jan;29(12):2881.