Please use this identifier to cite or link to this item: https://hdl.handle.net/11147/2846
Title: Investigation of the molecular and genetic response in enterocytes of duodenum during elevated intracellular glucose level
Other Titles: İnce bağırsak enterosit hücrelerinde artan hücre içi glikoz miktarında bağlı moleküler ve genetik yanıtın araştırılması
Authors: Boztepe, Tuğçe
Advisors: Güleç, Şükrü
Seyrantepe, Volkan
Keywords: Glucose
Obesity
Human intestinal epithelial
CaCo-2 Cells
Type 2 diabetes
Enterocyte cells
Publisher: Izmir Institute of Technology
Source: Boztepe, T. (2016). Investigation of the molecular and genetic response in enterocytes of duodenum during elevated intracellular glucose level. Unpublished master's thesis, İzmir Institute of Technology, İzmir, Turkey
Abstract: Glucose is one of the nutritional factor that involves in developing of obesity and type 2 diabetes in human. The studies indicated that enterocyte cells on intestine might play a role in dietary glucose sensing during obesity. Obese people are consumed high amount of dietary glucose and enterocyte cells consequently are exposed to high glucose. Thus, we aimed to find relevant physiological pathways and genome-wide mRNA expression profiles that can be regulated by glucose in fully differentiated human intestinal epithelial (CaCo-2). The cells were maintained two different glucose levels (5.5mM for control, 25mM for high glucose) at least three passages. The cells were grown on transwell system for 21 days to mimic human intestine system. Transepithelial electrical resistances (TEER) were measured to control monolayer formation and polarization. RNA isolation was performed and whole genome mRNA expression profile were determined following gene ontology analysis to find affected molecular pathways. Compared to control relative glucose level was found high in basolateral side of CaCo-2 cells that were under high glucose condition without effecting TEER. GLUT2, SGLT1, GLUT5 mRNA levels were significantly reduced during elevated glucose levels which is consistent with literature. Significant fold change analysis showed that 351 genes upregulated and 468 genes under high glucose condition. We found high glucose significantly leads changes of molecular pathways (downregulated; glycolysis and gluconeogenesis, adherens junction, fructose/mannose metabolism, pentose phosphate pathway and upregulated; protein export). These results provide us better understanding and open new window for glucose metabolism of enterocytes during obesity.
Glikoz, obezite ve tip 2 diyabet gelişimine neden olan beslenme faktörlerinden biridir. Araştırmalar gösteriyor ki ince bağırsaktaki enterosit hücreler obezitede glikozun hissedilmesinde rol oynayabilir. Obez insanlar yüksek oranda glikoz içeren besinler ile besleniyorlar ve bunun sonucunda enterosit hücreleri yüksek glikoza maruz kalıyor. Bu sebepten dolayı, farklılaşmış insan bağırsak hücresi CaCo-2 da glikoz ile bağlantılı fizyolojik yolakların ve tüm genom mRNA profilinin bulunması amaçlanmıştır. Hücreler iki farklı glikoz seviyesinde (5.5mM kontrol için, 25mM yüksek glikoz için) en az üç kere pasajlanmıştır. Hücreler transwell sistem üzerinde büyütülmüştür ve 21 gün sonra insan bağırsak sistemini taklit etmiştir. Hücrelerin oluşturdukları elektriksel direnç değeri, hücrelerin monolayer yapı ve polarize oluşumunu kontrol etmek için ölçülmüştür. Daha sonra RNA izolasyonu yapıldı ve tüm genom mRNA profili incelenip gen ontoloji analizine göre etkilenmiş metabolik yolaklar bulundu. GLUT2, SGLT1 ve GLUT5 mRNA seviyeleri yükselen glikoz seviyesi nedeniyle önemli derecede azaldı ki bu sonuçlar literatür ile de doğru orantılıdır. Yüksek glikoz ortamında 351 genin arttığı ve 468 genin de azaldığı görülmüştür. Yüksek glikozun önemli derece metabolik yolaklara etki ettiğini bulduk (azalanlar; glikoliz ve glikoneogenesis, fruktoz ve mannoz metabolizması, pentoz fosfat ve artan; protein export). Bu sonuçlar obezitede enterosit glikoz metabolizması için bize yeni pencereler açmayı sağlayacaktır.
Description: Thesis (Master)--Izmir Institute of Technology, Biotechnology, Izmir, 2016
Includes bibliographical references (leaves: 60-70)
Text in English; Abstract: Turkish and English
x, 70 leaves
URI: http://hdl.handle.net/11147/2846
Appears in Collections:Master Degree / Yüksek Lisans Tezleri

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