SİSPLATİNE BAĞLI OTOTOKSİSİTEDE DARBEPOETİNİN KORUYUCU ROLÜ

Year 2022, Volume: 36 Issue: 3, 309 - 320, 27.01.2023

Asuman Feda Bayrak , Yuksel Olgun , Hande Evin Eskicioğlu , Selen Kum Özşengezer , Efe Özgür Serinan , Hatice Efsun Kolatan , Hasan Oğuz Çetinayak , Günay Kırkım , Safiye Aktaş , Osman Yılmaz , Nur Olgun , Zekiye Altun

https://doi.org/10.18614/deutip.1231950

Abstract

Amaç: Darbepoetin-alfa (DPO) uygulamasının sisplatine bağlı ototoksik etkiyi önleyici rolü olup olmadığının in-vivo olarak değerlendirilmesi amaçlandı.
Materyal metod: Çalışmada 28 adet wistar albino rat kullanılarak 4 grup oluşturuldu. Grup 1 (intraperitoneal) IP salin verilen kontrol grubu, Grup 2 sisplatin (16mg/kg IP tek doz), Grup 3 DPO (25 μg/kg IP), Grup 4 sisplatin verilmeden 24 saat önce ve verildikten yarım saat sonra 25 μg/kg tek doz IP DPO uygulanan grup. Ratlara ajan uygulamalarından önce ve deneyin 7. gününde distorsiyon ürünü otoakustik emisyon (DPOAE) ve işitsel uyarılmış beyin sapı potansiyelleri (ABR) yapıldı. İşitme ölçümleri sonrasında ratlar sakrifiye edildi. Apoptotik hücre ölümü kulak dokusunda kaspaz-3, -8, -9 ve Nrf2 ve Hem oksijenaz-1 (HO-1), NQQ1, Glutatyon (GST) gibi Nrf2 ilişkili antioksidan ve iNOS, nNOS ekspresyon düzeyleri immünohistokimyasal yöntemle çalışıldı. Serum glutatyon (GSH) ve anti-inflamatuar TNF-α ve IL-1 protein düzeyleri ELISA kitleri kullanılarak belirlendi.
Bulgular: Sisplatin uygulaması ile işitme ölçümlerinde kontrole göre ototoksik etkileri gösteren tüm frekanslarda işitme kaybı saptandı. İmmunohistokimyasal doku düzeyinde gerçekleştirilen incelemelerde sisplatin uygulanan grupta iç kulakta strüktürel değişiklikler, beyin ve sinir dokularında nekroz ve nekroptoz saptandı. Darbepoetinin sisplatinin ortaya çıkardığı iç kulak ve beyin hasarlarını önleyici etkileri hem apoptotik protein ekspresyonları hem de oksidatif stres ile ilişkili belirteçler olan Nrf-2 ve ilişkili HO-1, NQO1, iNOS ve nNOS, GST ve anti-inflamatuar proteinleri üzerinden saptandı.
Sonuç: Bu çalışmada sisplatinin oluşturduğu ototoksisitede DPO’in koruyucu etkileri apoptozun azaltılması, antioksidan Nrf-2 ve hedef proteinler ve glutatyon düzeylerinin artışı yanında anti-inflamatuar proteinler üzerinden geliştiği gösterilmiştir.

Keywords

Darbepoetin, Cisplatin, Ototoxicity, Nrf2, antioxidant system

Supporting Institution

Dokuz Eylül Üniversitesi

Project Number

2016.KB.SAG.038

Thanks

Bu çalışma DEU Bilimsel Araştırma Projeleri Koordinasyon Birimi (Proje no: 2016.KB.SAG.038) tarafından desteklenmiştir.

PROTECTIVE ROLE OF DARBEPOETIN IN CISPLATIN-INDUCED OTOTOXICITY

Abstract

AIM: It was aimed to evaluate in-vivo whether darbepoetin-alpha (DPO) application has a preventive role in the cisplatin-induced ototoxic effect.
MATERIALS and METHODS: In the study, four groups were formed using 28 Wistar albino rats. Group 1 (intraperitoneal) IP saline given control group, Group 2 cisplatin (16mg/kg IP single dose), Group 3 DPO (25 μg/kg IP), and Group 4 is the group in which a single dose of 25 μg/kg IP DPO was administered 24 hours before and half an hour after cisplatin administration. Distortion product otoacoustic emission (DPOAE) and brainstem auditory evoked potentials (BAEP) were performed on rats before agent administration and on the 7th day of the experiment. After hearing measurements, the rats were sacrificed. Apoptotic cell death in ear tissue, caspase-3, -8, -9 expression, Inducible Nitric Oxide Synthase (iNOS), Neuronal Nitric Oxide Synthase (nNOS) expression levels, antioxidant Nrf2 (Nuclear factor (erythroid-derived 2)) - like 2) and related Heme oxygenase-1 (HO-1), NAD(P)H quinone oxidoreductase 1 (NQQ1) were studied by immunohistochemical method. Serum glutathione (GSH) and anti-inflammatory TNF-α and IL-1β protein levels were determined using ELISA kits.
RESULTS: The hearing loss was detected at all frequencies showing ototoxic effects compared to control in hearing measurements with cisplatin application. In the examinations performed at the immunohistochemical tissue level, structural changes in the inner ear, necrosis, and necroptosis in the brain and nerve tissues were detected in the cisplatin-administered group. The preventive effects of darbepoetin on the inner ear and brain damage induced by cisplatin are detected by apoptotic protein expressions and oxidative stress-related markers, iNOS and nNOS, Nrf-2 and related HO-1, NQO1, serum GSH, and TNF-α and IL-1β proteins.
CONCLUSION: In this study, the protective effects of DPO in ototoxicity caused by cisplatin were demonstrated by reducing apoptosis, increasing antioxidant Nrf-2 and target proteins and glutathione levels, and through anti-inflammatory proteins.

Keywords

Darbepoetin, Cisplatin, Ototoxicity, Nrf2, antioxidant system

Project Number

2016.KB.SAG.038

References

• 1. Olgun Y, Kırkım G, Kolatan E, Kıray M, Bagrıyanık A, Olgun A, et al. Friend or foe? Effect of oral resveratrol on cisplatin ototoxicity. Laryngoscope. 2014 Mar;124(3):760-6
• 2. Olgun Y, Kırkım G, Altun Z, Aktaş S, Kolatan E, Kıray M, et al. Protective Effect of Korean Red Ginseng on Cisplatin Ototoxicity: Is It Effective Enough? J Int Adv Otol. 2016 Aug;12(2):177-183.
• 3. Borse V, Al Aameri RFH, Sheehan K, Sheth S, Kaur T, Mukherjea D, et al. l. Epigallocatechin-3-gallate, a prototypic chemopreventative agent for protection against cisplatin-based ototoxicity. Cell Death Dis. 2017 Jul 13;8(7)
• 4. Monge AN, Gassmann M, Bodmer D. Erythropoietin but not VEGF has a protective effect on auditory hair cells in the inner ear. Cell. Mol. Life Sci. 2009; 66:3595–3
• 5. Messier AM, Ohls RK. Neuroprotective Effects of Erythropoiesis Stimulating Agents in Term and Preterm Neonates. Curr Opin Pediatr. 2014 April; 26(2): 139–145
• 6. Berkingali N, Warnecke A, Gomes P, Paasche G, Tack J, Lenarz T, et al. Neurite outgrowth on cultured spiral ganglion neurons induced by erythropoietin. Hearing Research 243 (2008) 121–126
• 7. Forbes CA, Worthy G, Harker J, Kleijnen J, Kutikova L, Zelek L, et al. Dose Efficiency of Erythropoiesis-Stimulating Agents for the Treatment of Patients With Chemotherapy-InducedAnemia: A Systematic Review. Clinical Therapeutics 2014; 36(4):594-610 8. Ehnert S, Freude T, Eicher C, Burkhar B, Martinez JJ, Neumann J, et al. Darbepoetin inhibits proliferation of hepatic cancer cells in the presence of TGF β. Arch Toxicol.2014; 88:89–96 9. Mann C, Lee JH, Liu J, Stammers AM, Sohn HM, Tetzlaff W, et al. Delayed treatment of spinal cord injury with erythropoietin or darbepoetin—A lack of neuroprotective efficacy in a contusion model f cord injury. Experimental Neurology.2008; 211:34–40
• 10. Seymen P, Aytac E, Esen F, Tel Ç, Demir F, Genç H, et al. Darbepoetin Alpha Ameliorates Neuronal Damage in a Rat Model of Acute Ethanol Intoxication. International Journal of Neuroscience.2013; 123, 99–103 11. Kertmen H, Gürer B, Yilmaz ER, Arikok AT, Kanat MA, Ergüder BI, et al. The comparative effects of recombinant human erythropoietin and darbepoetin-alpha on cerebral vasospasm following experimental subarachnoid hemorrhage in the rabbit. Acta Neurochir. 2014; 156:951–962
• 12. Gunes D, Kirkim G, Kolatan E, Güneri EA, Ozoğul C, Altun Z, et al. Evaluation of the effect of acetyl L-carnitine on experimental cisplatin ototoxicity and neurotoxicity. Chemotherapy. 2011; 57:186-194.
• 13. Choi DE, Jeong JY, Lim BJ, Lee KW, Shin YT, Na KR. Pretreatment with Darbepoetin Attenuates Renal Injury in a Rat Model of Cisplatin-Induced Nephrotoxicity. The Korean Journal of Internal Medicine Vol. 24, No. 3, September 2009
• 14. Somdaş MA, Güntürk İ, Balcıoğlu E, Avcı D, Yazıcı C, Özdamar S. Protective effect of N-acetylcysteine against cisplatin ototoxicity in rats: a study with hearing tests and scanning electron microscopy. 15. Kim SJ, Kwak HJ, Kim DS, Choi HM, Sim JE, Kim SH, et al. Protective mechanism of Korean Red Ginseng in cisplatin-induced ototoxicity through attenuation of nuclear factor-κB and caspase-1 activation. Mol Med Rep. 2015Jul;12(1):315-22.
• 16. Spasojevic SD, Stojanovic VD, Barisic NA, Doronjski AR,Zikic DR, Babovic SM. Neuroprotective effects of hypothermia and erythropoietin after perinatal asphyxia in newborn rats. J Matern Fetal Neonatal Med. 2013; 26(15): 1506–1509
• 17. Maxwell JR, Ohls RK. Update on Erythropoiesis-Stimulating Agents Administered to Neonates for Neuroprotection. Neoreviews. 2019 Nov;20(11):622-635
• 18. Cruz Navarro J, Pillai S, Ponce LL, Van M, Goodman JC, Robertson CS. Endothelial nitric oxide synthase mediates the cerebrovascular effects of erythropoietin in traumatic brain injury. Front Immunol. 2014 Oct 9;5:494
• 19. Estfanous RS, Elseady WS, Kabel AM, Abd Ellatif RA. Amelioration of Cisplatin-Induced Ototoxicity in Rats by L-arginine: The Role of Nitric Oxide, Transforming Growth Factor Beta 1 and Nrf2/HO-1 Pathway. Asian Pac J Cancer Prev. 2020 Jul 1;21(7):2155-2162.