Ekspresija prijenosnika uključenih u prijenos glukoze u gastrointestinalnom sustavu: SGLT1, GLUT2 i Na+/K+ ATP-aze u mišjem modelu upalnih bolesti crijeva

master's thesis

• Title (english): The expression of glucose transporters in the gastrointestinal system: SGLT1, GLUT2 and Na+/K+ ATP-ase in mouse inflammatory bowel disease model
• Author: Anesa Čajević Glojnarić
• Mentor: Lidija Bach-Rojecky (mentor)
• Committee member: Lidija Bach-Rojecky (predsjednik povjerenstva)
• Committee member: Milena Jadrijević-Mladar Takač (član povjerenstva)
• Committee member: Hrvoje Brzica (član povjerenstva)
• Granter: University of Zagreb; Faculty of Pharmacy and Biochemistry; (Department of pharmacology); Zagreb
• Defense date and country: 2021-02-10, Croatia
• Scientific / art field, discipline and subdiscipline: BIOMEDICINE AND HEALTHCARE; Pharmacy; Medical Biochemistry

Abstract

Iz pregleda literature jasno je vidljiva povezanost metabolizma ugljikohidrata, IBD-a i šećerne bolesti, kao i uloga prijenosnika monosaharida kod navedenih stanja. Cilj ovog rada je utvrditi ekspresiju prijenosnika uključenih u prijenos glukoze u gastro intestinalnom sustavu; SGLT1, GLUT2 i Na/K ATP-aze u mišjem DSS modelu koristeći Enzyme-Linked Immunosorbent Assay (ELISA) test. Promatrat će se i utjecaj farmakološkog standarda na ekspresiju navedenih transportera. Muški miševi C57BL/6 soja bili su podijeljeni u tri grupe: kontrolnu grupu nosača (0.5% metil celuloza), grupu koja je dobivala nosač i 3% DSS i grupu koja je uz DSS dobivala i ciklosporin A (80 mg/kg/PO/QD). Žrtvovanje je provedeno osmi dan i mjereni pokazatelji su bili – promjena tjelesne mase, ocjene konzistencije stolice i fekalnog krvarenja, indeks aktivnosti bolesti, duljina kolona, hematološki i histopatološki parametri te ELISA metodom koncentracije inzulina, serumskog amiloid A proteina, GLUT2, SGLT1 i Na/K ATP-aze. Za ispitivanje statističke značajnosti između grupa korišten je Mann – Whitney U test (p < 0,05). U grupi tretiranoj 3% DSS-om zabilježen je značajan porast ocjena konzistencija stolice, kolorektalnog krvarenja, indeksa aktivnosti bolesti te histološke Nishitani ocjene u odnosu na kontrolnu grupu, dok su ocjene u ciklosporinskoj grupi bile niže nego u DSS grupi. Duljina kolona bila je značajnije skraćena u DSS grupi nego u ciklosporinskoj. Hematološki parametri crvene krvne slike bili su sniženi, a ukupan broj leukocita kao i subpopulacija bio je povišen u DSS grupi. U DSS grupi pale su vrijednosti glukoze, a povećala se koncentracija inzulina. Nije primijećen porast ekspresije SGLT1, dok je GLUT2 u DSS grupi bio značajno viši u duodenumu, a niži u jejunumu, u usporedbi s kontrolom. Ekspresija GLUT2 u ciklosporinskoj grupi bila je viša no u DSS grupi. Značajno povećanje ekspresije Na/K ATP-aze zamijećeno je u kolonu i jejunumu životinja DSS grupe. Dobiveni rezultati podupiru dosadašnja saznanja o povezanosti ugljikohidrata i IBD-a, međutim potrebno je napraviti veći broj detaljnijih analiza kako bi se preciznije odredila uloga navedenih prijenosnika u DSS modelu, a tako i u IBD-u.

Abstract (english)

Available literature clearly shows the connection between the carbohydrate metabolism, inflammatory bowel disease (IBD) and diabetes, as well as the relevance of glucose transporters in these conditions. The aim of this work is to determine the expression of the transporters included in the transport of glucose across the intestinal barrier: Sodium Glucose transporter 1 (SGLT1), Glucose transporter 2 (GLUT2), and Sodium/pottasium ATPase (Na/K ATPase) in the mouse Dextrane Sodium Sulfate (DSS) model using Enzyme-Linked Immunosorbent Assay (ELISA). The effect of Cyclosporyne A, a pharmacological standard, on the expression of the listed transporters will be measured as well. Male C57BL6 mice were allocated in three groups: control vehicle (0.5% methyl cellulose), vehicle treated group challenged with 3% DSS and finally the group challenged with 3% DSS and treated with Cyclosporine A (80 mg/kg/PO/QD). During the course of the study body weight loss, stoll consistency and colorectal bleeding were measured and the sampling was performed on day 8 of the study. Colon length, haemathologic and histopathologic parameters were also measured. The concentrations of insulin and serum amiloid A protein in the serum, as well as the expression of GLUT2, SGLT1 and Na/K ATP-ase were measured by ELISA. Mann – Whitney U test (p < 0,05) was used to calculate statistical significance. The group treated with 3% DSS showed significant increase of stool consistency, colorectal bleeding, disease activity index and Nishitani scores when compared to the control group, while the group treated with Cyclosporine A showed significant reduction of these scores. Haematological parameters were decreased, while total leukocyte count, as well as subpopulation counts were increased in the DSS challenged group. The DSS group showed lower concentrations of glucose while the insulin concentration was increased. No significant change of the SGLT1 expression was measured in the small intestines, while GLUT2 showed increased and decreased expression in duodenum and jejunum, respectively, when compared to control group. Significant increase in the expression of Na/K ATP-ase was observed in jejunum and colon in the group of animals treated with DSS. These results support current knowledge regarding the link between the carbohydrates and IBD, however more detailed analyses are required in order to clearly describe the role of glucose transporters in the DSS model of IBD.

• Keywords: DSS kolitis; GLUT 2; SGLT1; Na+/K+ ATP-aza
• Keywords (english): DSS colitis; GLUT2; SGLT1; Na+/K+ ATP-ase
• Language: croatian
• URN:NBN: urn:nbn:hr:163:509148
• Study programme: Title: Medical biochemistry; Study programme type: university; Study level: integrated undergraduate and graduate; Academic / professional title: magistar/magistra medicinske biokemije i laboratorijske medicine (magistar/magistra medicinske biokemije i laboratorijske medicine)
• Type of resource: Text
• File origin: Born digital
• Access conditions: Open access
• Created on: 2021-11-16 16:28:43