Title Uloga izraženosti MMP-9 i COX-2 proteina u analizi karotidnog aterosklerotskog plaka u bolesnika s karotidnom endarterektomijom Title (english) The expression of MMP-9 and COX-2 proteins in the analysis of carotid atherosclerotic plaques in patients with carotid endarterectomy Author Davorin Šef Mentor Anita Škrtić (mentor)Mentor Vinko Vidjak (komentor)Committee member Tomislav Meštrović (predsjednik povjerenstva)Committee member Branko Malojčić (član povjerenstva)Committee member Majda Vučić (član povjerenstva)Granter University of Zagreb Defense date and country 2022-04-11, Croatia Scientific / art field, BIOMEDICINE AND HEALTHCARE Universal decimal classificationUDC ) 616 - Pathology. Clinical medicine Abstract Predoperativnom radiološkom dijagnostikom (ultrazvuk/Color Doppler – UZV/CD, magnetska rezonancija - MR, višeslojna kompjutorizirana tomografija - MSCT) mogu se analizirati komponente karotidnog aterosklerotskog plaka (KAP). Imunohistokemijskim bojenjem endarterektomiranih karotidnih plakova može se procijeniti izraženost proteina matriks metaloproteinaze 9 (MMP-9) i ciklooksigenaze 2 (COX-2) koji imaju ulogu u remodeliranju aterosklerotskog plaka.
Cilj istraživanja bio je usporediti radiološke, patohistološke i imunohistokemijske karakteristike KAP-a u simptomatskih i asimptomatskih bolesnika s indikacijom za karotidnu endarterektomiju (CEA). Hipoteza istraživanja bila je da su radiološka obilježja nestabilnosti karotidnog plaka analiziranog predoperativnom neinvazivnom radiološkom dijagnostikom (UZV/CD, MR i/ili MSCT) značajno povezana s izraženošću MMP-9 i COX-2 proteina u KAP-u i da su oni visoko zastupljeni u bolesnika s nestabilnim plakom.
Istraživanje je bilo prospektivno kohortno i prijavljeno je u registar ISRCTN (ID ISRCTN46536832). U istraživanju je učinjena predoperativna radiološka dijagnostika karotidnih arterija, patohistološka analiza te analiza imunohistokemijskog bojenja endarterektomiranih karotidnih plakova.
U istraživanje je bio uključen 31 bolesnik s indikacijom za endarterektomiju unutarnje karotidne arterije (CEA) od čega je 13 simptomatskih i 18 asimptomatskih bolesnika medijana dobi od 70 (64 - 75) godina, među kojima je bilo 18 muškaraca (58,1 %) i 13 žena (41,9 %). U skupini simptomatskih bolesnika, njih 8 (62 %) je predoperativno imalo tranzitornu ishemičku ataku (TIA) dok je preostalih 5 (38 %) bolesnika imalo cerebrovaskularni inzult (CVI). Medijan trajanja hospitalizacije je bio 5 (3 - 6) dana. Nije bilo perioperativne smrtnosti. MR i MSCT dijagnostikom su u 12 od 13 (92,3 %) simptomatskih bolesnika dijagnosticirani nestabilni plakovi (P = 0,020). Među bolesnicima s radiološki dijagnosticiranim nestabilnim plakom je nađena značajno veća mikrovaskularna gustoća unutar plaka (P = 0,049), dok je u simptomatskih bolesnika uočena značajno veća zastupljenost veće masne jezgre (P = 0,009). Semikvantitativnom analizom je utvrđeno da su bolesnici s nestabilnim plakovima imali značajno veću izraženost te intenzitet bojenja COX-2 (P = 0,038, odnosno P = 0,010) u CD68 PG-M1 pozitivnim makrofagima. Dodatnom analizom korelacija i ROC analizom, potvrđene su značajne pozitivne, srednje jake korelacije većeg intenziteta bojenja COX-2 u CD68 PG-M1 pozitivnim makrofagima unutar KAP-a s prisutnošću predoperativnih simptoma. Uočili smo značajno veću izraženost MMP-9 u CD68 PG-M1 pozitivnim makrofagima u simptomatskih bolesnika (P = 0,020), ali izraženost MMP-9 nije bila značajno različita pri usporedbi radiološki nestabilnih i stabilnih plakova.
Predoperativne neinvazivne radiološke dijagnostičke metode mogu poboljšati probir bolesnika s nestabilnim plakom u kojih je indicirana CEA. Naši rezultati pokazuju da je COX-2 povezan s radiološki dijagnosticiranom nestabilnošću karotidnog plaka. MMP-9 bi mogao biti povezan s povećanim rizikom od cerebrovaskularnih komplikacija.
Abstract (english) Components of carotid atherosclerotic plaque can be analysed by non-invasive preoperative radiological imaging (ultrasound/color doppler - USS/CD, magnetic resonance imaging - MRI and multi-slice computed tomography - MSCT) The expression of matrix metalloproteinase 9 (MMP-9) and cyclooxygenase-2 (COX-2) proteins, that have a role in remodelling of atherosclerotic plaques, can be analysed immunohistochemically. The aim of this study was to analyse the radiological, pathohistological and immunohistochemical characteristics of carotid atherosclerotic plaques (CAP) in symptomatic and asymptomatic patients with an indication for carotid endarterectomy (CEA). We hypothesised that radiological features of unstable plaque, analyzed by preoperative noninvasive radiological diagnostics (USS/CD, MR and/or MSCT), can be associated with the expression of MMP-9 and COX-2 proteins in CAP and that they are highly represented in patients with unstable plaque. This prospective observational study has been registered at the ISRCTN registry (ID ISRCTN46536832). Preoperative radiological imaging, as well as postoperative pathohistological and immunohistochemical analyses were performed.
We included 31 patients (42% symptomatic and 58% asymptomatic, median age of 70 (64-75) years), of which 18 males (58.1%) and 13 females (41.9%), with an indication for CEA. In the group of symptomatic patients, 8 (62%) had transient ischemic attack preoperatively, while the other 5 (38%) patients had a cerebrovascular stroke. Median duration of hospitalization was 5 (3-6) days. There was no perioperative mortality. Unstable plaques were detected in 12/13 (92.3%, P=0.020) symptomatic patients using MR and/or MSCT. A significantly higher microvascular density within the plaque was found (P=0.049) among patients with radiologically unstable plaque, while a large lipid core was found significantly more often (P=0.011) among symptomatic patients. The expression and intensity of COX-2 staining in CD68 PG-M1 positive macrophages by semiquantitative analysis were significantly higher among unstable when compared to stable (P=0.038 and P=0.010, respectively). Additional correlation and ROC analysis confirmed significant positive correlations of the high intensity COX-2 staining in CD68 PG-M1 positive macrophages within CAP, and the presence of preoperative symptoms. We observed significantly higher expression of MMP-9 in CD68 PG-M1 positive macrophages among symptomatic patients (P=0.020), but there was no significantly difference when comparing the expression of MMP-9 between radiologically unstable and stable plaques.
Advanced non-invasive multimodality preoperative imaging could help in improving identification of patients with unstable plaque that have an indication for CEA. Our results suggest that COX-2 is associated with the unstable plaque detected with preoperative imaging. MMP-9 could be associated with an increased risk of cerebrovascular complications.
Keywords
Matriks metaloproteinaza 9
Ciklooksigenaza 2
Aterosklerotski plak
Karcinoembrionalni antigen
Ishemijski napad
Endarterektomija
Mikrovaskularna gustoća
Moždani udar
Magnetska rezonancija
Keywords (english)
Matrix Metalloproteinase 9
Cyclooxygenase 2
Atherosclerotic Plaque
Carcinoembryonic Antigen
Ischemic Attack
Endarterectomy
Microvascular Density
Stroke
Magnetic Resonance Imaging
Language croatian URN:NBN urn:nbn:hr:105:167314 Study programme Title: Biomedicine and Health Sciences Type of resource Text Extent 129 str. File origin Born digital Access conditions Open access Terms of use Created on 2022-04-26 07:24:37
