Evaluation of MMP-9 and iNOS expressions in sheep with encephalitic listeriosis

Öz

This study aimed to correlate Matrix Metalloproteinase-9 and iNOS expressions with the severity of histopathological findings in tissue samples taken from sheep with encephalitic listeriosis. Thus, the role of these molecules in the pathogenesis of the disease can be elucidated. After systemic necropsy, tissue samples of adult sheeps with meningoencephalitis were investigated by the culture, histopathological and immunohistochemical methods in the presence of Listeria spp. isolation from tissues was performed in accordance with the USDA-FSIS method with some modifications. Tissue samples were fixed in a 10% buffered formaldehyde solution. Following routine procedures, tissue sections at 5 μm were stained with Hematoxylin and Eosin, investigated under light microscope and photographed. Immunohistochemical staining was performed on the tissues using the avidin-biotin immune peroxidase complex method. Listeria spp. were obtained in 20 (83.3%) of 24 tissue samples with the presence of bright grey-black centred smooth colonies on Listeria Selective Agar and identified as Listeria monocytogenes through the phenotypically supportive tests. Liquefaction necrosis, purulent meningoencephalitis, perivascular cuffing, microabscesses and glial nodules were the most important histopathological findings. MMP-9 immunpositive reactions were observed in the cytoplasm of microglial cells and neurons in areas where inflammatory and necrotic areas are concentrated in medulla oblongata and pons. In perivascular cuffing areas, immune reactions in endothelial cells were detected. We detected iNOS positive reactions in the medulla oblangata and pons, especially in inflammatory cells in the microabscesses. Consequently, a positive correlation (p < 0.05) was found between MMP-9 expression and the severity of histopathological findings in sheep with encephalitic listeriosis. expression. In addition, we found that iNOS expression increased in parallel with the increase in MMP-9

Anahtar Kelimeler

• culture
• encephalitic listeriosis
• histopathology
• immunohistochemistry
• mmp-9

Ensefalitik listeriyozisli koyunlarda MMP-9 ve iNOS ekspresyonunun değerlendirilmesi

Öz

Bu çalışmada ensefalitik listeriyozisli koyunlardan alınan doku örneklerinde gözlenen histopatolojik bulguların şiddeti ile Matriks metalloproteinaz-9 ve iNOS ekspresyonlarını korele etmeyi amaçladık.Böylece bu moleküllerin hastalığın patogenezindeki rolü açıklanabilecektir. Sistemik nekropsi sonrası meningoensefalitli erişkin koyunlardan alınan doku örnekleri Listeria spp. varlığı için kültür, histopatolojik ve immunohistokimyasal olarak incelendi. Dokulardan Listeria spp. izolasyonu bazı modifikasyonlarla USDA-FSIS yöntemine uygun olarak gerçekleştirildi. Doku örnekleri %10’luk tamponlu formaldehit solüsyonunda tespit edildi. Rutin işlemlerden sonra 5 μm kalınlığındaki kesitler Hematoksilen&Eozin ile boyandı, ışık mikroskobu altında incelendi ve fotoğraflandı. Dokulara immunohistokimyasal boya olarak avidin-biotin immunperoksidaz kompleks metodu uygulandı. Listeria spp. Listeria Selective Agar üzerinde parlak gri-siyah merkezli pürüzsüz koloniler bulunan 24 doku örneğinden 20'sinde (% 83.3) elde edildi ve fenotipik olarak destekleyici testler yoluyla Listeria monocytogenes olarak tanımlandı. Likefaksiyon nekrozu, purulent meningoensefalitis, perivasküler hücre infiltrasyonu, mikroapseler ve glial nodüller en önemli histopatolojik bulgulardı. MMP-9 immunpozitif reaksiyonları yangının ve nekrozun yoğun olduğu alanlardaki mikroglial hücreler ve nöronların sitoplazmasında gözlemledik. Perivasküler hücre infiltrasyonu alanlarında, endotelyal hücrelerde de immun reaksiyonu saptadık. iNOS pozitif reaksiyonları özellikle medulla oblongata ve pons bölgesinde yer alan mikroapselerdeki yangısal hücrelerde tespit ettik. Sonuç olarak ensefalitik listeriyozisli koyunlarda MMP-9 ekspresyonu ile histopatolojik bulguların şiddeti arasında pozitif bir korelasyon tespit ettik (p<0.05). Buna ek olarak MMP-9 ekspresyonundaki artışa paralel olarak iNOS ekspresyonun da artış gösterdiğini ortaya koyduk.

Anahtar Kelimeler

• ensefalitik listeriyozis
• histopatoloji
• immunohistokimya
• kültür
• mmp-9

Kaynakça

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