Title Izražaj apoptotskih i proliferacijskih čimbenika u želučanoj sluznici oboljelih od sistemske skleroze
Title (english) Expression of apoptotic and proliferative factors in the gastric mucosa of patients with systemic sclerosis
Author Katarina Borić
Mentor Snježana Mardešić (mentor)
Committee member Dijana Perković (predsjednik povjerenstva)
Committee member Snježana Tomić (član povjerenstva)
Committee member Ozren Polašek (član povjerenstva)
Granter University of Split School of Medicine Split
Defense date and country 2021, Croatia
Scientific / art field, discipline and subdiscipline BIOMEDICINE AND HEALTHCARE Clinical Medical Sciences Internal Medicine
Universal decimal classification (UDC ) 616 - Pathology. Clinical medicine
Abstract U istraživanju „Izražaj apoptotskih i proliferacijskih čimbenika u želučanoj sluznici oboljelih
od sistemske skleroze“ histološki su analizirane razlike u proliferaciji, apoptozi i profibrotičkim
čimbenicima u uzorcima želučane sluznice oboljelih od sistemske skleroze (SSc) i kontrolne
skupine. U istraživanju je sudjelovao 15 bolesnika sa SSc i 5 kontrolnih ispitanika bez SSc.
Bolesnici su sistematizirani s obzirom na oblik i težinu kliničke slike sistemske skleroze. Pet
bolesnika je klasificirano u skupinu s ograničenom kožnom SSc (lcSSc) i blagom bolešću, pet
s umjerenom difuznom i pet s teškom difuznom kožnom SSc (dcSSc).
Uzorci tkiva su imunohistokemijski obojeni protutijelom Ki-67 (proliferacija), kaspazom-3
(apoptoza), sindekanom-1 (epitelni biljeg), CTGF (profibrotički biljeg) i α-SMA (biljeg
miofibroblasta). Istovremena prisutnost (kolokalizacija) Ki-67 pozitivnih stanica s biljegom za
miofibroblaste (α-SMA) određena je upotrebom dvostruke imunofluorescentne metode.
Postotak reaktivnih stanica analiziran je primjenom Kruskal-Wallis testa i Dunn-ovog post hoc
testa, a P<0.05 se smatrao statistički značajnim.
Analiza je otkrila da je postotak proliferirajućih (Ki-67 pozitivnih) stanica bio je najveći u
uzorcima bolesnika s umjerenom dcSSc (3% u površinskim i 4,6% u dubljim djelovima
želučane sluznice, koji je pao na 1% u uzorcima bolesnika s teškom dcSSc). Postotak stanica
pozitivnih na α-aktin glatkih mišića (α-SMA) iznosio je 5% u uzorcima kontrola, 9% u
površinskim djelovima želučane sluznice bolesnika s lcSSc, dok je u dubljem sloju porastao na
19% u uzorcima bolesnika s umjerenom i 14% s teškom dcSSc, što ukazuje na povećanu
populaciju i aktivnost miofibroblasta. Pozitivne apoptotske stanice na kaspazu-3 iznosile su
1,5-2% u kontrolnim uzorcima, 8% u površinskim i 6% u dubljim dijelovima želučane sluznice
bolesnika s umjerenom dcSSc. U uzorcima bolesnika s teškom dcSSc, apoptoza je zahvatila
50% površinskih epitelnih i žljezdanih stanica i 30% dubljih žlijezda, a korelirala je s
povećanom ekspresijom CTGF i fibrozom te smanjenom ekspresijom sindekana-1.
Ovi podaci pokazuju da je umjerena dcSSc proliferacijski najaktivniji oblik SSc. Tešku dcSSc
karakteriziraju taloženje kolagena, defekti površinskog epitela, opsežna apoptoza i snižena
proliferacija, atrofija i gubitak funkcije.
Abstract (english) In the study "Expression of apoptotic and proliferative factors in the gastric mucosa of patients
with systemic sclerosis" differences in proliferation, apoptosis and profibrotic factors were
histologically analyzed in samples of gastric mucosa in patients with systemic sclerosis (SSc)
and control group. The study included 15 patients with SSc and 5 control subjects without SSc.
Patients were systematized with respect to the type and severity of systemic sclerosis. Five
patients were classified into the group with limited cutaneous SSc (lcSSc) and mild disease,
five with moderate diffuse, and five with severe diffuse cutaneous SSc (dcSSc).
Tissue samples were immunohistochemically stained with Ki-67 antibody (proliferation),
caspase-3 (apoptosis), syndecan-1 (epithelial marker), CTGF (profibrotic marker), and α-SMA
(myofibroblast marker). The simultaneous presence (colocalization) of Ki-67 positive cells with
α-SMA was determined using double immunofluorescence method. The percentage of reactive
cells was analyzed by a Kruskal-Wallis test and the Dunn post hoc test. P<0.05 was considered
statistically significant.
The analysis revealed that the percentage of proliferating (Ki-67 positive) cells was highest in
samples of patients with moderate dcSSc (3% in superficial and 4.6% in deeper parts of the
gastric mucosa, which dropped to 1% in samples of patients with severe dcSSc). The percentage
of α-smooth muscle actin (α-SMA) positive cells was 5% in control samples, 9% in superficial
gastric mucosa of patients with lcSSc, while in the deeper layer it increased to 19% in samples
of patients with moderate and 14% with severe dcSSc, indicating increased myofibroblast
population and activity. Apoptotic cells positive on caspase-3 account for 1.5–2% in control
samples, 8% in superficial, and 6% in deeper parts of the gastric mucosa of patients with
moderate dcSSc. In samples of patients with severe dcSSc, apoptosis involved 50% of
superficial epithelial and glandular cells and 30% of deeper glands, and correlated with
increased CTGF expression and fibrosis, and decreased syndecan-1 expression.
This data show that moderate dcSSc is the most proliferatively active form of SSc. Severe
dcSSc is characterized by collagen deposition, surface epithelial defects, extensive apoptosis
and decreased proliferation, atrophy and loss of function.
Keywords
sistemska sklerodermija
želučana sluznica
apoptoza
proliferacija stanica
faktor rasta vezivnog tkiva
Keywords (english)
Systemic Scleroderma
Gastric Mucosa
Apoptosis
Cell Proliferation
Connective Tissue Growth Factor
Language croatian
URN:NBN urn:nbn:hr:171:648753
Study programme Title: Translational Research in Biomedicine - TRIBE Study programme type: university Study level: postgraduate Academic / professional title: doktor/doktorica znanosti, interdisciplinarna područja znanosti (doktor/doktorica znanosti, interdisciplinarna područja znanosti)
Type of resource Text
File origin Born digital
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Created on 2023-05-03 13:14:11