Aktivnost glukuronidacijskog sustava u metabolički važnim tkivima miša

Autor: Đolonga, Petar
Přispěvatelé: Terzić, Janoš, Markotić, Anita, Zekić Tomaš, Sandra, Vilović, Marino
Jazyk: chorvatština
Rok vydání: 2023
Předmět:
Popis: Ciljevi istraživanja: Međusobno usporediti bazalni izražaj gena Ugt1a u metabolički važnim tkivima miša. ii Utvrditi promjenu izražaj gena Ugt1a u jetri, bubregu i tankom crijevu nakon akutnog tretmana BBN-om. iii Odrediti utječe li redukcija mikrobiote antibioticima na izražaj gena sustava Ugt u mokraćnom mjehuru miševa tretiranih BBN-om. iv Odrediti postoji li promijenjen izražaj pojedinačnih izoformi gena sustava Ugt u tumorima mokraćnog mjehura miševa. Materijali i metode: Eksperiment je izveden u Laboratoriju za istraživanje raka pri Katedri za imunologiju i medicinsku genetiku Medicinskog fakulteta Sveučilišta u Splitu. Od sedam miševa soja C57BL/6J četiri su uzimala BBN u vodi za piće (pokusna grupa), a tri su pila vodu (kontrolna grupa). Metodom qRT-PCR kvantificiran je izražaj gena Ugt1a u jetri, bubregu, duodenumu, jejunumu i bedrenom. Nadalje, provjerena je aktivnost gena glukuronidacijskog sustava analizirajući ranije prikupljene podatke RNA sekvenciranja tumorskog tkiva kao i zdravog tkiva mokraćnog mjehura miša nakon antibiotske redukcije mikrobiote i tretmana BBN-om. Rezultati: Jetra, duodenum, jejunum i bubreg imaju značajno veću ekspresiju, glavnog glukuronidacijskog gena Ugt1a nego bedreni mišić. U odnosu na jetru, samo je bubreg pokazao značajno veći izražaj gena Ugt1a. Nije pronađena značajna promjena izražaja gena Ugt1a nakon tretmana miševa BBN-om. U tumorskom tkivu mokraćnog mjehura miševa primijećena je značajno niža ekspresija šest izoformi gena sustava Ugt, i to: Ugt1a6a, Ugt1a6b, Ugt1a7c, Ugt2b1, Ugt2b34 i Ugt2b35. U mokraćnom mjehuru miševa s antibiotski reduciranom mikrobiotom i tretiranih BBN-om primijećen je statistički značajno manji izražaj gena Ugt1a7c. Zaključak: Rezultati ovog istraživanja ukazuju da jetra nije jedini organ sa značajnom glukuronidacijskom sposobnošću jer je visoka razina izražaja gena Ugt1a pronađena u bubregu i tankom crijevu. Konzumacija BBN-a nije značajno promijenila izražaj gena Ugt1a u metabolički relevantnim tkivima miša. Mikrobiota bi, također, mogla imati važnu ulogu u regulaciji izražaja gena iz sustava Ugt u mokraćnom mjehuru. Smanjen izražaj gena za enzime glukuronidacijskog sustava u tumorskom tkivu mokraćnog mjehura ukazuje da je smanjenje glukuronidacijske sposobnosti možda povezano s urotelnom karcinogenezom.
Objectives: i To compare the basal expression of the Ugt1a gene in metabolically important mouse tissues. ii To determine the change in the expression of the Ugt1a gene in the liver, kidney, and small intestine after acute treatment with BBN. iii To determine whether the reduction of microbiota with antibiotics affects the expression of the Ugt system in the urinary bladder of mice treated with BBN. iv To determine whether there is an altered expression of individual isoforms of the Ugt genes in mice bladder tumors. Materials and methods: The experiment was conducted in the Cancer Research Laboratory at the Department of Immunology and Medical Genetics, University of Split School of Medicine. Of the seven C57BL/6J mice, four took BBN in drinking water (experimental group), and three drank water (control group). The expression of the Ugt1a gene in the liver, kidney, duodenum, jejunum, and femoral muscle was quantified using the qRT-PCR method. Furthermore, the activity of the glucuronidation system genes was analyzed using previously collected RNA sequencing data of urinary bladder tumor tissue as well as mouse urinary bladder tissue after antibiotic reduction of microbiota and two-week BBN treatment. Results: Liver, duodenum, jejunum, and kidney have significantly higher expression of the main glucuronidation gene Ugt1a compared to the skeletal muscle. Compared to the liver, only the kidney showed a significantly higher expression of the Ugt1a gene. No significant change in Ugt1a gene expression was found after treatment of mice with BBN. In the murine urinary bladder tumor tissue, a significantly lower expression of six Ugt isoforms was observed, namely: Ugt1a6a, Ugt1a6b, Ugt1a7c, Ugt2b1, Ugt2b34 and Ugt2b35. A statistically significant lower expression of the Ugt1a7c gene was observed in the urinary bladder of mice with antibiotic-reduced microbiota and treated with BBN. Conclusion: The results of this study indicate that the liver is not the only organ with significant glucuronidation capacity, as a high level of Ugt1a gene expression was found in the kidney and small intestine. BBN consumption did not significantly change the expression of the Ugt1a gene in metabolically relevant mouse tissues. The microbiota could also play an important role in the regulation of urinary bladder Ugt genes expression. Decreased expression of genes for enzymes of the glucuronidation system in urinary bladder tumor tissue indicates that a possible decrease in glucuronidation capacity may be related to the urothelial carcinogenesis.
Databáze: OpenAIRE
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