Analysis of apoptosis of kidney tissue by the tunel method and histomorphological changes in rabbit kidney model due to unilateral supravesical obstruction

Öz

Aim: There is a need for effective, cheap, and fast method to detect apoptosis. In some studies, we see that newer, more difficult, expensive, or less effective methods are used. We wanted to show that the TUNEL method has serious advantages and can still be used alone. In this study, it was aimed to investigate whether there is a significant difference in the number of apoptotic cells in partial obstruction (PO) and complete obstruction (CO) by using terminal deoxytransferase-mediated bio-dUTP nick and labeling (TUNEL) method in renal tissue. We also evaluated histopathological changes after renal obstruction. Methods: In this study, 29 rabbits were used. Supravesical obstruction was created in 24 rabbits. Five rabbits were used as the control group. Twelve kidneys were examined after creating unilateral partial obstruction and 12, after the creation of unilateral complete obstruction. Histomorphological changes in kidney tissues in routine Haemetoxylene-Eosine (HE) preparations and apoptosis in preparations obtained by TUNEL method were examined. Results: Apoptotic cells were observed especially in the tubules by the TUNEL method. The average number of apoptotic cells in CO and PO groups were 190.66 and 40.58, respectively. In the CO group, the number of apoptotic cells was significantly higher than that in the PO group (P<0.001). Interstitial fibrosis, chronic inflammatory infiltration, tubular destruction (vacuolar changes, cystic and atrophic tubules) were observed in both groups. These changes were more limited, and mild, in the PO group, and severe and widespread in the CO group. Conclusion: TUNEL method is one of the highly effective methods in detecting apoptosis. It was observed that apoptosis and pathological changes developing in the kidney tissue after complete obstruction were more severe and widespread.

Anahtar Kelimeler

• Renal obstruction
• Apoptosis
• TUNEL method

Tavşan böbrek modelinde tek taraflı supravezikal tıkanıklığa bağlı böbrek dokusunda oluşan apoptozisin tunel yöntemiyle analizi ve histomorfolojik değişiklikler

Öz

Amaç: Literatürde, bazı çalışmalarda, daha yeni, ama pratikte değerlendirme güçlükleri olan, pahalı veya etkinliği az yöntemlerin kullanıldığını görüyoruz. Terminal deoxytransferase-mediated bio-dUTP nick and labeling (TUNEL) yönteminin ciddi avantajları olduğunu ve halen tek başına kullanılabileceğini göstermek istedik. Bu nedenle, deneysel olarak tavşan üreterinde kısmi ve tam obstrüksüyon oluşturulmasından sonra böbrek dokusunda gelişen apopitozisin şiddetinin, TUNEL yöntemi kullanılarak belirlenmesi, gruplar arasında anlamlı bir fark olup olmadığı ve yanısıra oluşan histopatolojik değişiklikleri değerlendirme amaçlandı. Yöntemler: Mesane üstü seviyesinde üreter geçişinin kısmi ve tam olarak engellenmesi yoluyla-12 tam obstrüksiyon, 12 kısmi obstrüksiyon grubu-, 5 taneside kontrol grubu olmak üzere- toplam 29 adet tavşan böbreği kullanıldı. Böbrek dokusunda gelişen histomorfolojik değişiklikler Hematoksilen-Eozin (HE) preperatlarda, apoptozis ise TUNEL yöntemiyle hazırlanan preperatlarda mikroskopik olarak incelendi. Obstrüksiyon sonrası oluşan apopitozisin tam ve kısmi obstrüksiyon grupları arasında istatistiksel olarak anlamlı farklılığı olup olmadığı araştırıldı. Bulgular: Apopitotik hücreler, TUNEL yöntemiyle net olarak gösterildi. Apopitozisin, özellikle tübül epitelinde yoğunlaştığı görüldü. Tam obstrüksiyon grubunda, ortalama apopitotik hücre sayısı 190,66 ve kısmi obstriksüyon grubunda ise 40,58 idi. Tam obstrüksiyon grubunda apopitotik hücre sayısı kısmi obstrüksiyon grubuna göre anlamlı derecede yüksekti (P<0,001). 2 grupta da interstisyel fibrozis, kronik iltihabi infiltrasyon, tübüler yıkım (vakuoler değişiklikler, kistik ve atrofik tübüller) gözlendi. Bu değişiklikler kısmi tıkanma grubunda daha sınırlı, hafif, tam tıkanma grubunda şiddetli ve yaygın idi. Sonuç: TUNEL yönteminin apopitozisi belirlemede etkinliği yüksek yöntemlerden biri olduğu gösterildi. Tam tıkanma sonrası böbrek dokusunda gelişen apopitozisin ve patolojik değişikliklerin çok daha şiddetli ve yaygın olduğu görüldü.

Anahtar Kelimeler

• Renal obstrüksiyon
• Apopitozis
• TUNEL yöntemi

Kaynakça

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