Sağlıklı ve Diabet oluşturulmuş farelerin böbrek dokusunda katalaz enziminin RT-PCR ile gen ve immunohistokimyasal olarak protein ekspresyonu

Bu çalışma; deneysel Diabetes Mellitus (DM) oluşturulan farelerin böbrek dokusundaki katalaz enzimi (CAT) gen ekspresyonu düzeyinin, bu enzimin dokudaki lokalizasyonunun ve diabetli hayvanların böbrek dokusunda oluşan histolojik değişikliklerin belirlenmesi amacıyla yapıldı. Çalışmada kullanılan 36 adet Swiss albino fare, deneme (n=15), sham (n=15) ve kontrol (n=6) gruplarına ayrıldı. Deneme grubuna, 100 mg/kg dozda Streptozotosin (STZ) intraperitoneal (İP) yolla uygulandı. Deneme grubunda kan glikoz düzeyi 200 mg/dl’nin üzerinde olan fareler diabetik kabul edilerek çalışmaya alındı. CAT mRNA ekspresyon düzeyi Reverse Transkripsiyon-Polimeraz Zincir Reaksiyonuyla (RT-PCR), bu enzimin böbrek dokusundaki lokalizasyonu ise immunohistokimyasal yöntemle belirlendi. CAT gen ekpresyonu ve enzimatik aktivitesinin deneme grubunda sham ve kontrol gruplarına göre daha düşük olduğu tespit edildi (P

Anahtar Kelimeler:

enzimler, kan şekeri, enzim aktivitesi, katalaz, hayvan modelleri, ters transkriptaz, fareler, böbrekler, immünohistokimya, genler, gen ifadesi

The gene expression profile by RT-PCR and immunohistochemical expression pattern of Catalase in the kidney tissue of both healthy and diabetic Mice

The aim of this study was to determine the gene expression pattern of catalase by RT-PCR technique, its tissue localization by immunohistochemistry method and the histological changes of DM kidney by routine histological techniques. In this study, 36 swiss albino mice were used and the animals were allocated into 3 groups as experimental (n=15), sham (n=15) and control (n=6) were used. STZ (100 mg/kg) was applied intraperitoneally (IP) to experimental group. Mice in which blood glucose level were above 200 mg/dl were included into DM group and considered as experimental group for the present study. Catalase gene expression level was determined by means of RT-PCR. Tissue localization of catalase enzyme was analysed by immunohistochemical method. Gene expression and enzyme activity of catalase from experimental group were found to be lower than those of the sham and control groups (P<0.05). Catalase staining was mainly observed in the renal cortex of all groups, whereas its reaction was relatively weaker in DM group than that of the other groups. Glycogen level was decreased in the proximal tubuls and hydropic degeneration was found especially in distal tubuls of the diabetic kidney. Result of the present study might make a contribution to future studies in the field of DM aiming to establish a better understanding of the relationship between antioxidants and DM.

Keywords:

enzymes, blood sugar, enzyme activity, catalase, animal models, reverse transcriptase, mice, kidneys, immunohistochemistry, genes, gene expression,

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