Yenidoğan streptozotosin diyabet modelinde pankreatik beta hücre rejenerasyonunu kontrol eden genlerin ekspresyonu

Bu çalışmanın amacı, yenidoğan streptozotosin (yd-STZ) diyabet modelinde, erken dönemlerde beta hücre rejenerasyonunu takip etmek amacıyla, insülin, pankreatik duedonal homeobox gen-1 (pdx-1) ve sinaptofisin gen ekspresyonlarını hem adacık içi hem de adacık dışı hücrelerdeki dağılımlarını tespit etmektir. Ayrıca STZ uygulaması ile bozulan adacık düzeninin yeniden yapılanmasında, endokrin hücrelerin rolünü araştırmaktır. Çalışmada, her birinde 7 adet Wistar albino yenidoğan sıçanlardan oluşan 8 grup oluşturuldu. 4 gruba doğumun 2. günü 100 mg/kg tek doz STZ (yd2-STZ) uygulaması yapıldı ve gruplar sırasıyla doğumun 3., 5., 7. ve 10. günlerinde sağlıklı kontrol grupları ile birlikte dekapite edildiler. Alınan pankreas dokuları % 10’luk formolde tespit edilerek parafine gömüldü. Doku kesitlerine insülin probu kullanılarak in situ hibridizasyon yöntemi, ayrıca insülin, glukagon, somatostatin, sinaptofisin, pdx-1 antikorları kullanılarak immünohistokimyasal boyama yöntemi uygulandı. STZ uygulanan gruplarda kontrol gruplarına göre adacıkların küçüldüğü, Yd2- STZ diyabetik gruplarda insülin mRNA sinyallerini içeren hücrelerin kapladığı alanın, insülin immün pozitif hücrelerin kapladığı alandan daha az olduğu görüldü. Yd2-STZ gruplarında adacık içerisindeki immün pozitif pdx-1 hücrelerinin kontrol gruplara göre azaldığı ayrıca tüm deney gruplarında adacıklardaki tüm endokrin hücrelerin sinaptofisin ekspresyonu yaptıkları tespit edildi. STZ uygulanan gruplarda ekzokrin doku ve kanal epiteli içerisinde çok sayıda sinaptofisin immün pozitif hücrelere rastlandı. yd2-STZ modelinde, STZ’nin neden olduğu beta hücre hasarının en fazla görüldüğü günün yd-2 STZ 5 günlük grup olduğu ve 5. günden başlayarak 7 ila 10. günler arasında hızlı bir rejenerasyonun meydana geldiği, sinaptofisin ve pdx-1’in birlikte, olası öncül hücrelerin endokrin hücreye dönüşümünü tespit etmek için iyi birer belirteç olacakları sonucuna varıldı.

The expression of genes that regulat pancreatic beta cell regeneration in neonatal streptozotocin diabetic model

The aims of this study are to determine distribution of gene expression of insulin, pdx-1 and synaptophisin in both intra and extra islet cells, to determine the role of the endocrine cells in the remodelling of islets that was damaged with STZ treatment in the early stages of the neonatal STZ (n-STZ) diabetic model. In this study 8 groups each containing 7 neonatal Wistar albino rats were established. On the second day after birth 100mg/kg STZ (n-STZ) was given four groups. Each group was decapitated on the 3rd, 5th, 7th and 10th day respectively together with the healthy control groups. Pancreas tissue was fixed in % 10 formaline and embedded in paraffin. Digoxigenin labeled insulin was used for in situ hybridization. Insulin, glucagon, somatostatin, synaptophisin and pdx-1 antibodies were used for immunohistochemistry on serial parafine sections. Islet sizes of STZ treated groups were smaller than control groups, while the area containing insulin mRNA signal positive cells in n-STZ diabetic group were smaller than controls. Although pdx-1 immunopositive cells were decreased in STZ diabetic groups compared to control groups. Numerous synaptophysin positive cells were detected in the lining of duct epithelium as well as exocrine tissue in the STZ diabetic groups. Beta cell destrucition was in the highest level in n-STZ 5 days group, and an effective beta cell regeneration occured between 7 th and 10 th day of the experiment. Synapthophysin and pdx-1 may be a useful marker in the detection of precursor cells transforming to β cells.

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