Kezban YILDIZ DALGINLI, Melek ÖZTÜRKLER, Hatice BEŞEREN, Yasemen ADALI, Onur ATAKİSİ

Ratlarda Sisplatin Kaynaklı Akciğer Toksisitesi Üzerine Oksidatif/Nitrozatif Stres Parametrelerinin ve Histopatolojik, İmmünohistokimyasal Etkilerin Değerlendirilmesi

Bu çalışmanın ana odak noktası, sisplatin kaynaklı akciğer toksisitesinde oksidatif/nitrozatif stres ve antioksidan etkiler ile immünohistokimyasal etkileri araştırmaktır. Çalışmada 2 aylık 12 Sprague Dawley erkek rat kontrol (n=6) ve sisplatin (n=6) olmak üzere iki gruba ayrıldı. Kontrol grubuna izotonik solüsyon, sisplatin grubuna sisplatin 10 mg/kg tek doz intraperitoneal uygulandı. Alınan akciğer dokularında redükte glutatyon (GSH), malondialdehit (MDA) ve nitrik oksit (NO) seviyeleri spektrofotometrik yöntemle belirlendi. Akciğer dokularından parafin bloklar yapıldı ve hematoksilen-eozin (HE) boyama ile boyandı. İmmünohistokimyasal olarak p53, CD3, CD20, Bcl-2 ve Ki67 değerlendirildi. Sıçanların akciğer dokusunda tek başına sisplatin uygulamasının MDA ve GSH değerleri üzerine etkisinin olmadığı, NO düzeylerinin anlamlı olarak arttığı bulundu (P<0.005). Akciğer dokusunun histopatolojik değerlendirmesinde HE boyama ile konjesyon-kanama bulguları, yoğun inflamasyon alanları, bronş ve bronşiyol çevresinde lenfoid foliküller görüldü. Alveoller arasında düşük yoğunluklu ödem ile hava yollarında kan-fibrin ve enflamatuar hücreler ve fibroblastlardan oluşan konsantrik fibröz ve fibrinöz tıkaçlar gözlendi. Reaktif pan B hücre belirteçleri CD20, interstisyel bileşende T-hücre belirteci CD3 ve subepitelyal hücrelerde desmin immünohistokimyasal boyama ile pozitif boyanırken, bronşiyoller ve alveolar kanallara uygulanan reaktif germinal merkezler Bcl-2 ve p53 immünohistokimyasal olarak negatif boyandı. Ayrıca Ki67 immünohistokimyasal boyama ile düşük yoğunluklu nükleer boyanma saptandı. Sonuç olarak NO düzeyinde anlamlı artış ve immünohistokimyasal olarak yoğun inflamasyon, lenfoid foliküller, fibröz ve fibrnöz tıkaçlar sisplatin kaynaklı akciğer hasarı başlangıcının bir ifadesidir.

Anahtar Kelimeler:

Sisplatin, oksidatif/nitrozatif stres, histopatoloji, immünohistokimya, rat

Evaluation of Oxidative/Nitrosative Stress Parameters and Histopathological, Immunohistochemical Effects on Cisplatin-Induced Lung Toxicity in Rats

The main focus of this study is to investigate oxidative/nitrosative stress and antioxidant effects and immunohistochemical effects in cisplatin-induced lung toxicity. In the study, 12 male Sprague Dawley rats, 2 months old, were divided into two groups: control (n=6) and cisplatin (n=6). Isotonic solution was administered to control and cisplatin 10 mg/kg single dose intraperitoneal to cisplatin group. Reducte glutathione (GSH), malondialdehyde (MDA) and nitric oxide (NO) levels were determined by spectrophotometricmethod in the lung tissues taken. Paraffin blocks were made from lung tissues and stained with hematoxylin-eosin (HE) staining. Immunohistochemically, p53, CD3, CD20, Bcl-2 ve Ki67 were evaluated. It was found that cisplatin administration alone had no effect on MDA and GSH values in the lung tissue of rats, and NO levels were significantly increased (P<0.005). In the histopathological evaluation of the lung tissue, congestion-bleeding findings, intense inflammation areas, lymphoid follicles around the bronchi and bronchioles were seen with HE staining. Concentric fibrous and fibrinous plugs consisting of blood-fibrin and inflammatory cells and fibroblasts in the airways were observed, with low-density edema between the alveoli. Reactive pan B cell markers CD20, T-cell marker CD3 in the interstitial component and desmin in sub-epithelial cells were stainedpositively by immunohistochemical staining, while reactive germinal centers Bcl-2 and p53 applied to the bronchioles and alveolarducts were immunohistochemically stained negative. In addition, low-intensity nuclear staining was found with Ki67 immunohistochemical staining.In conclusion, significant increase in NO level and immunohistochemically intense inflammation, lymphoid follicles, fibrous and fibrinous plugs are an expression of the onset of cisplatin-induced lung injury..

Keywords:

Cisplatin, oxidative/nitrosative stress, histopathology, immunohistochemical, rat,

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