Title Vino kao modulator rane upalne faze cijeljenja akutnog infarkta miokarda na štakorskom modelu
Title (english) Wine as a modulator of the early inflammatory phase of acute myocardial infarction healing in a rat model
Author Nikola Ključević
Mentor Ivica Grković (mentor)
Committee member Valdi Pešutić-Pisac (predsjednik povjerenstva)
Committee member Damir Fabijanić (član povjerenstva)
Committee member Luka Zaputović (član povjerenstva)
Granter University of Split School of Medicine Split
Defense date and country 2019, Croatia
Scientific / art field, discipline and subdiscipline BIOMEDICINE AND HEALTHCARE Basic Medical Sciences Pharmacology
Universal decimal classification (UDC ) 615 - Pharmacology. Therapeutics. Toxicology
Abstract Naš je zadatak bio „titrirati“ ranu (upalnu fazu) cijeljenja akutnog infarkta miokarda
kvantitativnom i kvalitativnom analizom infarciranog i rubnog ishemičnog, kao i
neinfarciranog dijela miokarda s obzirom na izražaj upalnih markera MMP-2, MMP-9, NFκB
i TGFβ1 i upalnih stanica (neutrofili i monociti/makrofazi) koji se javljaju neposredno nakon
infarkta i tijekom prvih 24 sata od inicijalnog događaja na štakorskom modelu. Saznanja koja
imamo o upalnoj fazi cijeljenja infarkta miokarda stavili smo u odnos s umjerenom
konzumacijom bijelog vina te ispitali blagotvorni učinka prekondicioniranja standardnom i
maceriranom Graševinom.
Životinje su randomizirane u skupine i 28 dana su konzumirale ili vodu ili bijelo vino
(standardno i macerirano bijelo vino), nakon čega su podvrgnute operativnom zahvatu
podvezivanja predenjeg silaznog ogranka lijeve koronarne arterije izazivajući infarkt
miokarda. Nakon 24 sata inkubacije životinje su žrtvovane, srca eksplantirana, te su se
hisotološki odredile 3 reprezentativne zone, (zona infarkta, kontrolna zona, peri-infarktna
zona), u kojima se mjerio izražaja pozitivnog signala gore navedenih upalnih markera i broj
pozitivnih upalnih stanica pomoću protutijela na MPO (neutrofili) i CD68
monociti/makrofazi).
Nađen je značajno manji izražaj MMP-9 markera u svim reprezentativnim zonama kod
životinja koje su pile vino, bez obzira na sadržaj fenola (p<0.001). Ipak, atenuacija signala je
bila značajno slabija kod skupine koja je konzumirala macerirano vino. Slični trendovi su
primijećeni i za marker NFκB. Nije bilo razlike u imunoreaktivnosti MMP-2 među
skupinama, osim u peri-infarktnoj zoni, gdje je uočena atenuacija signala kod skupine koja je
konzumirala standardno vino (p<0.004). Nije bilo razlike u ekspresiji markera MMP-2
između skupine koja je pila vodu i skupine koja je pila macerirano vino u peri-infarktnoj zoni.
Razlika nije pronađena u ispitivanim skupinama za marker TGF-β1 ni u jednoj od
reprezentativnih zona. Pronašli smo smanjen izražaj pozitivnih stanica na MPO i CD68 u
peri-infarktnoj zoni životinja koje su pile vino (p<0.001). U sham grupi opisan je
subepikardijalni „upalni prsten/ sloj“ stanica.
Znanstvena vrijednost i doprinos ove doktorske disertacije je u prvi put eksperimentalno
opisanom kardioprotektivnom učinku konzumacije bijelog vina. Pokazali smo da umjerena
konzumacija bijelog vina kroz razdoblje od mjesec dana prije infarkta značajno pridonosi
smanjenju izražaja ključnih upalnih markera u tkivu štakora, što bi moglo povoljno utjecati na
tijek i oporavak nakon oboljenja.
Abstract (english) Two main points were adressed in this disertation. Effects of white wine and macerated white
wine consumption on the expression of inflammatory markers/mediators (MMP-2, MMP-9,
NF-κB and TGF-β1) and on inflammatory cells infiltration (neutrofils and
monocytes/macrophages) in myocardial tissue after experimentally induced permanent
myocardial ischemia were investigated. Male Sprague-Dawley rats were given either a
combination of different white wines or water only, for 28 days. After coronary ligation,
animals were left to survive for 24 hours. Three representative areas: infarct/ischemic, periinfarct/
border zone, and control/non-ischemic zones were analyzed for the expression of
immunoreactivity by measuring the threshold area % of signal density for inflammatory
markers. Same areas were analysed for total number of MPO (neutrophils) and CD68
(moncytes-macrophages) positive cells per area of view.
For MMP-9, significantly smaller expression was found in all 3 zones of wine drinking
animals (P < 0.001). There was no difference in MMP-2 immunoreactivity between two
groups, except in peri-infarct zones, where the signal was significantly decreased (P < 0.001).
The same pattern of expression was found for the NF-κB p65 signal, although no differences
between experimental groups were observed for TGF-β1. There was no difference between
polyphenol- rich white wine and standard white wine consumption for any of the
abovementioned markers.
Smaller expression for both MPO and CD68 was found in all three peri-infarct zones of wine
drinking animals (p<0.001). There was no difference in expression of leukocyte markers
between animals drinking standard and polyphenol-rich white wine, although for CD68, a
non-significant attenuation was noticed. In sham animals, a sub-epicardial MPO/CD68
immunoreactive ‘inflammatory ring’ is described. Standard white wine consumption caused
attenuation of expression of MPO but not of CD68 in these animals.
Main highlights of our investigation are that white wine consumption, regardless of its
polyphenol content, decreases the expression of the investigated inflammatory
markers/mediators in the peri-infarct zone, suggesting its significant modulatory potential. For
MMP-9 and MMP-2, expression was similar to the effect of postischemic reperfusion. No
effect on TGF-β1 was observed, highlighting its role in being the masterswitch, changing
from the inflammatory to the proliferative stage of infarct healing. Finally, we conclude that
white wine consumption positively modulates peri-infarct inflammatory infiltration.
Keywords
infarkt miokarda
modeli bolesti na životinjama
medijatori upale
infiltracija neutrofila
štakori Sprague-Dawley
vino
Keywords (english)
Myocardial Infarction
Animal Disease Models
Inflammation Mediators
Neutrophil Infiltration
Sprague-Dawley Rats
Wine
Language croatian
URN:NBN urn:nbn:hr:171:361495
Project Number: IP-2013-11-8652 Title: Biološki učinci vina: utjecaj vinifikacijske tehnologije, dealkoholizacije i starenja vina Title: Biological effects of wine: the influence of vinification technology, dealcoholization and aging of wine Acronym: BioWine Leader: Mladen Boban Jurisdiction: Croatia Funder: HRZZ Funding stream: IP
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)
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Created on 2023-05-10 07:02:33