MUSA KARAMAN, HASAN ÖZEN, KADİR ÖZCAN, Mehmet TUZCU

Immunohistochemical demonstration of nitrosative tissue damage in copper ınduced liver toxicity in Japanese quails

Bakırın fazlası çeşitli türlerde karaciğer toksikasyonuna neden olabilir. Bu çalışmada, bıldırcınlarda deneysel kronik bakır toksikasyonunda, karaciğerde oluşan nitrozatif doku hasarı ve apoptotik hücre ölümü immunohistopatolojik metodlarla incelendi. Her birinde 10 adet bir günlük bıldırcın bulunan kontrol, 1, 2 ve 3. gruplar, 6 hafta süre ile sırasıyla 0, 100, 250 ve 500 ppm düzeylerinde bakır sülfat içeren yemle beslendiler. Beslenme dönemi sonrasında, hayvanlara ötanazi uygulanarak karaciğerleri alındı ve histopatolojik yöntemler için rutin şekilde işlendi. İndüklenebilir nitrik oksit sentaz (iNOS) ve nitrotirozin için indirekt avidin-biotin peroksidaz yöntemi uygulandı. Apoptotik hücre ölümü in situ TUNEL boyama yöntemi uygulanarak araştırıldı. Mikroskobik düzeyde karaciğerde sadece 2. grupta hafif, 3. grupta orta derecede dejeneratif değişiklikler tespit edildi. İmmunohistokimyasal boyamalarda her iki antikor için de kontrol grubunda hafif derecede immun tepkime tespit edildi. Artan bakır konsantrasyonuna bağlı olarak immun tepkimelerde artma gözlendi. Sadece 3. grupta ve az sayıda olmak üzere apoptotik hepatositler mevcuttu. Elde edilen bu bulgular, bakır ile oluşturulan karaciğer toksikasyonunda, nitrik oksitin oksidatif doku hasarında rol oynadığını göstermektedir.

Anahtar Kelimeler:

toksik maddeler, dokular, bıldırcın, bakır, programlanmış hücre ölümü, zarar, karaciğer, immünohistokimya, toksikoloji, toksisite

Bıldırcınlarda bakır ile oluşturulan karaciğer toksikasyonunda nitrozatif doku hasarının immunohistokimyasal yöntemlerle araştırılması

Excess copper may cause liver toxicosis in various species. In this study, nitrosative tissue damage was investigated in an experimental chronic copper toxicity model in Japanese quails by immuno-histopathological means. Four groups (control, 1st, 2nd and 3rd), each comprised of 10 one-day-old chicks, were fed for 6 weeks with rations containing none, 100, 250 and 500 ppm copper sulfate, respectively. At the end of the feeding period, animals were euthanized, liver tissues were collected and processed routinely for histopathologic applications. Immunohistochemical staining was performed for inducible nitric oxide synthase (iNOS) and nitrotyrosine antibodies with indirect avidin-biotin peroxidase technique. Apoptotic cell death was investigated with in situ TUNEL staining. Microscopically, there were only mild and moderate degenerative changes in the liver of the chicks of 2nd and 3rd groups, respectively. In immunohistochemical staining, little immunoreactivity was detected in the control group for both antibodies. Immunoreactivities were gradually stronger with the increasing copper concentration. A few apoptotic hepatocytes were present only in the 3rd group. These findings suggest that nitric oxide play a role in the oxidative tissue damage of copper induced liver toxicity.

Keywords:

toxic substances, tissues, quails, copper, apoptosis, damage, liver, immunohistochemistry, toxicology, toxicity,

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