Immunohistochemical Investigation of Oxidative Stress-induced DNA Damage and Lipid Peroxidation in Bovine Papillomas and Fibropapillomas

Abstract

In this study, it was aimed to evaluate the expressions of 8-OHdG and MDA immunohistochemically in order to determine the oxidative stress-induced DNA damage and lipid peroxidation in 16 papillomas and 14 fibropapillomas from 30 cattle brought to our department between 2013-2020. Biopsy samples taken after surgery were fixed in buffered 10% formaldehyde solution. Sections of 5 μm thickness were taken from the paraffin blocks prepared after routine tissue follow-up procedures and Hematoxylin & Eosin staining was applied to the sections in order to detect histopathological changes. Avidin Biotin Peroxidase method was used for immunohistochemical staining. In papilloma cases, severe hyperkeratosis, retepects extending from the epidermis to the dermis, spongiosis and balloon-like degeneration in squamous epithelial cells, basophilic inclusion bodies in granular cells, ulcerations in the epidermis layer, hemorrhagic areas and an increase in kerato hyaline granules were observed. In addition to these findings, dense connective tissue increases were detected in fibropapilloma cases. In immunohistochemical evaluations, positive reactions for BPV were observed in the nuclei of cells in stratum granulosum. 8-OHdG positive reactions were detected in the cytoplasm and nucleus of the epidermal cells in papilloma cases, whereas in cases of fibropapilloma, reactions in these cells were observed in the cytoplasm of fibrocytes and fibroblasts in the dermis. Membranous MDA positive reactions were observed in the epidermal cells in papilloma cases, whereas MDA expressions were detected in the cytoplasm of fibrocytes and fibroblasts in the dermis of fibropapilloma cases. As a result of the literature reviews, no study data was found in which DNA damage due to oxidative stres and lipid peroxidation was detected by means of 8-OHdG and MDA expressions in papilloma and fibropapilloma cases of cattle, it is thought that the findings obtained in this study will contribute to the literature. In addition, it has been concluded that oxidative stress plays an important role in the pathogenesis of this tumor.

Keywords

• DNA damage
• oxidative stress
• papilloma
• lipid peroxidation

Sığır Papillom ve Fibropapillomlarda Oksidatif Stres Kaynaklı DNA Hasarı ve Lipid Peroksidasyonun İmmunohistokimyasal Olarak Araştırılması

Abstract

Bu çalışmada 2013-2020 yılları arasında anabilim dalımıza getirilen toplamda 30 adet sığırlara ait 16 adet papillom ve 14 adet fibropapillom örneğinde oksidatif stres kaynaklı DNA hasarı ve lipid peroksidasyonu belirlemek amacıyla immunohistokimyasal olarak 8-OHdG ve MDA ekspresyonlarının değerlendirilmesi amaçlandı. Cerrahi operasyon sonrası alınan biyopsi örnekleri tamponlu %10’luk formaldehit solüsyonunda fikze edildi. Rutin doku takip işlemleri sonrası hazırlanan parafin bloklardan 5 μm kalınlığında kesitler alındı ve histopatolojik değişikliklerin saptanabilmesi amacıyla kesitlere Hematoksilen & Eozin boyaması uygulandı. İmmunohistokimyasal boyamalarda Avidin Biotin Peroksidaz metodu uygulandı. Papillom vakalarında şiddetli hiperkeratoz, epidermisten dermise doğru uzanan retepektler, skuamöz epitel hücrelerinde spongiyozis ve balonumsu dejenerasyon, granüler hücrelerde bazofilik inklüzyon cisimcikleri, epidermis katmanında ülserasyonlar ve kanama alanları ile kerato hiyalin granüllerinde artış gözlendi. Fibropapillom vakalarında ise bu bulgulara ek olarak girdap tarzında bağ doku artışları tespit edildi. İmmunohistokimyasal değerlendirmelerde ise BPV pozitif reaksiyonlar stratum granulozumdaki hücrelerin çekirdeğinde gözlendi. 8-OHdG pozitif reaksiyonlar papillom vakalarında epidermal hücrelerin sitoplazmasında ve çekirdeğinde saptanırken, fibropapillom vakalarında ise bu hücrelerdeki reaksiyonlara ek olarak dermisteki fibrosit ve fibroblastların sitoplazmasında rastlanıldı. Papilloma olgularında epidermal hücrelerde membranöz MDA pozitif reaksiyonlar görülürken, fibropapilloma vakalarının dermisinde fibrosit ve fibroblastların sitoplazmasında MDA ekspresyonları tespit edildi. Yapılan literatür taramaları sonucunda sığırlara ait papillom ve fibropapillom vakalarında oksidatif strese bağlı DNA hasarı ile lipid peroksidasyonun 8-OHdG ve MDA ekspresyonları vasıtasıyla tespit edildiği herhangi bir çalışma verisine rastlanmamış olup ve bu yönüyle bu çalışmada elden edilen bulguların literatüre katkı sunacağı düşünülmektedir. Bunlara ek olarak oksidatif stresin bu tümörün patogenezinde önemli bir rol oynadığı kanaatine varılmıştır.

Keywords

• dna hasarı
• oksidatif stres
• papillom
• lipid peroksidasyon

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