Title UČINAK NEDOSTATKA TREFOIL
FAKTOR 3 PROTEINA NA JETRU I
HIPOKAMPUS MIŠEVA IZLOŽENIH
KRATKOTRAJNOM TRETMANU
VISOKOMASNOM HRANOM
Title (english) EFFECT OF TREFOIL FACTOR 3
PROTEIN DEFICIENCY ON
LIVER AND HIPPOCAMPUS OF
MICE ON SHORT TERM HIGH
FAT DIET TREATMENT
Author Kate Šešelja
Mentor Mirela Baus Lončar (mentor)
Committee member Ivana Ratkaj (predsjednik povjerenstva)
Committee member Igor Jurak (član povjerenstva)
Committee member Bojan Polić (član povjerenstva)
Granter University of Rijeka (Faculty of Biotechnology and Drug Development) Rijeka
Defense date and country 2023-06-20, Croatia
Scientific / art field, discipline and subdiscipline BIOTECHNICAL SCIENCES Biotechnology
Universal decimal classification (UDC ) 577 - Biochemistry. Molecular biology. Biophysics
Abstract Trefoil faktor protein 3 (Tff3) je mali peptid primarno eksprimiran u vrčastim stanicama probavila
gdje sudjeluje u zaštiti epitela od raznolikih štetnih utjecaja. Tff3 je prisutan i u jetri i u mozgu te
novija istraživanja ukazuju na potencijalno važnu ulogu u metaboličkim i neurodegenerativnim
poremećajima. Metaboličke bolesti kao što je Tip 2 dijabetes, i neurodegenerativne bolesti
uključujući Alzheimerovu bolest, dijele brojne patofiziološke poveznice. Proučavanje poveznica
u ranim fazama bolesti, prije uznapredovalih simptoma, bi moglo pridonijeti boljem razumijevanju
patogeneze ovih kompleksnih poremećaja i uspješnijem razvoju djelotvornih terapija. Stoga je cilj
ovog doktorskog rada bio istražiti ulogu Tff3 u jetri i hipokampusu u počecima aktivacije
patoloških procesa navedenih poremećaja uslijed kratkotrajnog tretmana visokomasnom hranom.
U tu svrhu je razvijen novi kongenični Tff3-/- mišji soj na C57BL/6N genskoj podlozi bez dodatnih
relevantnih metaboličkih mutacija. Zatim su novo-razvijeni Tff3-/- miševi oba spola te
odgovarajuće divlji tip kontrole hranjene visokomasnom hranom kroz 9 tjedana. Ispitan je
generalni metabolički status životinja, analizirana je ekspresija markera relevantnih
patofizioloških puteva u jetri te markera neurogeneze i neuroinflamacije u hipokampusu. Tff3-/-
životinje su imale statistički značajno manje mase i uočeno je smanjeno nakupljanje masti u jetri
u odnosu na WT kontrole što ukazuje na zaštitni učinak nedostatka Tff3. U jetri Tff3-/- mužjaka
je uočena značajno snižena razinu gena Il1α i Cxcr7, dok je u jetri Tff3-/- ženki povećana razina
Irs2, Atf4 gena te Ybx1 i Anp32a proteina. U hipokampusu Tff3-/- mužjaka snižena je razina
markera nezrelih neurona (Dcx) te markera mikroglija (Iba1), dok Tff3-/- ženke imaju povećanu
razinu Dcx te povećanu razinu markera zrelih neurona (NeuN). Analiza genske ekspresije markera
neurogeneze ukazuje na izraženije razlike u hipokampusu Tff3-/- ženki. U jetri i hipokampusu
životinja izloženih standardnoj hrani nisu detektirane statistički značajne razlike navedenih
promijenjenih markera što ukazuje kako se uočene promjene javljaju uslijed stresa izazvanog
visokomasnom hranom. Ovo istraživanje pruža novi uvid u uloge Tff3 proteina u jetri i
hipokampusu te pruža smjer za daljnja ispitivanja sa ciljem otkrivanja točnih mehanizama
djelovanja te potencijalno identifikacije novih terapijskih meta metaboličkih/neurodegenerativnih
poremećaja. Dodatno, ukazuje na potrebu za uključivanjem oba spola u buduća istraživanja s
obzirom da se uočen fenotip Tff3-/- mužjaka i Tff3-/- ženki bitno razlikuje.
Abstract (english) Trefoil factor protein 3 (Tff3) is a small peptide expressed mainly in goblet cells with a role
in healing and protection of epithelium. Recent research suggests that the Tff3 protein also
plays an important role in metabolic and neurodegenerative diseases. Metabolic diseases, such
as Type 2 diabetes and neurodegenerative diseases, such as Alzheimer's disease, share many
pathophysiological links. Studying these links in the early stages, before the onset of advanced
symptoms, could contribute to a better understanding of the pathogenesis of these complex
disorders and to the successful development of effective therapies. Therefore, the aim of this
doctoral thesis was to investigate the role of Tff3 in the liver and the hippocampus in the early
stages of pathological processes activation due to short-term treatment with a high-fat diet.
First, a new congenic Tff3-/- mouse strain was developed on the C57BL/6N genetic
background without additional relevant metabolic mutations. Subsequently, the newly
developed Tff3-/- mice of both sexes and corresponding wild-type controls were fed a highfat diet for 9 weeks. We examined the general metabolic status of the animals and analyzed
the expression of markers of relevant pathophysiological pathways in the liver and in the
hippocampus. Tff3-/- animals had significantly lower weight and reduced fat accumulation in
the liver compared with WT controls, suggesting a protective effect of Tff3 deficiency. The
results of molecular analysis showed decreased levels of the Il1α and Cxcr7 genes in the liver
of Tff3-/- males, whereas the levels of the Irs2 and Atf4 and Ybx1 and Anp32a were increased
in Tff3-/- females. In the hippocampus of Tff3-/- males, the protein level of the marker of
immature neurons (Dcx) and the marker of microglia (Iba1) were decreased, whereas levels
of Dcx and the marker of mature neurons (NeuN) were increased in Tff3-/- females. Analysis
of neurogenesis markers shows more pronounced effect in the hippocampus of Tff3-/- females.
No statistically significant differences in the aforementioned altered markers were detected in
both liver and hippocampus of the animals exposed to a standard diet, suggesting that the
observed changes are a consequence of stress induced by high-fat diet. This research provides
new insights into the role of Tff3 protein in the liver and the hippocampus and provides
direction for further investigations aimed at uncovering the precise molecular mechanisms. In
addition, it highlights the need to include both sexes in future research, as the observed
phenotype differs significantly by sex.
Keywords
Trefoil faktor 3
tretman visokomasnom hranom
metabolizam
neurogeneza
neuroinflamacija
Keywords (english)
Trefoil factor 3
high fat diet
metabolism
neurogenesis
neuroinflammation
Language croatian
URN:NBN urn:nbn:hr:193:885505
Promotion 2023
Study programme Title: Medicinal chemistry Study programme type: university Study level: postgraduate Academic / professional title: doktor/doktorica znanosti, interdisciplinarna područja znanosti, polje biotehnologija u biomedicini (doktor/doktorica znanosti, interdisciplinarna područja znanosti, polje biotehnologija u biomedicini)
Type of resource Text
File origin Born digital
Access conditions Open access
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Created on 2023-09-22 12:39:22