Title UTJECAJ PRIJEOPERACIJSKE PERORALNE PREHRANE NA RANI IMUNOLOŠKI ODGOVOR U BOLESNIKA S KORONARNOM BOLESTI SRCA LIJEČENIH KIRURŠKOM REVASKULARIZACIJOM MIOKARDA Title (english) INFLUENCE OF PREOPERATIVE PERORAL NUTRITION ON EARLY IMMUNE RESPONSE ON PATIENTS WITH CORONARY ARTERY DISEASE WHO UNDERWENT ON SURGICAL CORONARY REVASCULARIZATION Author Jadranko Sokolić Mentor Vlatka Sotošek Tokmadžić (mentor)Committee member Gordana Laškarin (predsjednik povjerenstva)Committee member Višnja Majerić-Kogler (član povjerenstva)Committee member Igor Medved (član povjerenstva)Granter University of Rijeka Defense date and country 2018, Croatia Scientific / art field, BIOMEDICINE AND HEALTHCARE Universal decimal classificationUDC ) 61 - Medical sciences ThesaurusMESH - Medical Subject Headings )
Coronary Artery Disease
Myocardial Revascularization
Preoperative Care
Perforin
Immunity, Cellular
Abstract Cilj istraživanja
Kirurška revaskularizacija miokarda značajno utječe na promjenu imunoloških parametara, osobito stanične imunosti, što može dovesti do slabljenja funkcije imunološkog sustava u cijelosti. Nepoznati su mehanizmi koji dovode do neravnoteže i slabljenja imunološkog sustava te utječe li i na koji način prijeoperacijsko hranjenje na imunološka zbivanja u bolesnika s koronarnom bolešću srca liječenih kirurškom revaskularizacijom miokarda. Cilj ovog rada bio je istražiti promjene udjela leukocitnih subpopulacija, izražaj citotoksične molekule perforina u pojedinim limfocitnim subpopulacijama, stupanj citotoksične aktivnosti stanica NK i izražavanje pojedinih citokina (interferona-gama: IFN, interleukin (IL)-15, IL-18) u krvi bolesnika s koronarnom bolešću srca liječenih kirurškom revaskularizacijom miokarda te istražiti kako prijeoperacijska peroralna prehrana utječe na navedene imunološke parametre.
Ispitanici, materijal i metode
Ovo prospektivno istraživanje je obuhvatilo bolesnike s koronarnom bolešću srca, koji su liječeni kirurškom revaskularizacijom miokarda. U studiji je ukupno sudjelovalo 80 bolesnika, podijeljenih u četiri grupe. Bolesnici iz Grupe 1 liječeni su kirurškom revaskularizacijom miokarda (CABG od eng. Coronary Artery Bypass Grafting) uz upotrebu stroja za vantjelesni krvotok (tzv."on-pump" CABG), a nisu bili hranjeni prijeoperacijski. U Grupi 2 bili su bolesnici liječeni s on-pump CABG koji su bili hranjeni prijeoperacijski. U Grupi 3 bili su bolesnici s CABG bez stroja za vantjelesni krvotok (off-pump CABG, OPCABG) a koji nisu bili hranjeni, dok su u Grupi 4 bili OPCABG bolesnici koji su uzimali hranu prijeoperacijski.
Svakom bolesniku uključenom u studiju uzeto je 20 ml heparinizirane periferne venske krvi neposredno prije kirurškog zahvata (T1) te nakon 24 (T2) i 72 (T3) sata po završetku operacije. Uzorci su centrifugirani na gradijentu gustoće i iz njih su izolirane mononuklearne stanice kojima su istodobno označeni antigeni korištenjem florokromnih konjugiranih monoklonalnih antitijela. Tim postupkom detektirane su leukocitne populacije (limfocite T, stanice NK, stanice NKT) i njihove subpopulacije. Izdvojenim, označenim stanicama metodom direktne imunofluorescencije unutar stanice obilježavan je perforin i analiziran protočnom citometrijom.
Za određivanje koncentracije IFNγ, IL-15 i IL-18 u plazmi bolesnika korišten je ELISA test (eng. enzyme linked immunosobrent assay).
Pomoću pozitivne magnetske separacije izdvojene su iz periferne krvi stanice NK i potom je dvosatnim testom citotoksičnosti i protočnom citometrijom određivana njihova citotoksična aktivnost.
Rezultati
Dvadeset i četiri sata nakon kirurškog zahvatauočen je statistički značajnosmanjenje učestalosti CD3+ i CD3+CD4+ stanica te udjela perforin pozitivnih i perforin+CD3+CD8+ stanica u bolesnika liječenih s on-pump CABG operacijom u usporedbi s bolesnicima liječenih off-pump CABG operacijom. Udio T regulacijskih stanica je statistički značajno porastao u bolesnika liječenih on-pump CABG operacijom u usporedbi s bolesnicima liječenih off-pump CABG operacijom 24 sata nakon operacije.
Udio IL-15+CD3+, IL-15+CD56+ te IL-15+CD3+CD56+ stanica bio je statistički značajno niži u bolesnika liječenih off-pump CABG operacijom u usporedbi s bolesnicima liječenih on-pump CABG operacijom 24 sata nakon oepracije. Istovremeno, nisu zabilježene značajne razlike u u udjelu IFNγ+ stanica unutar i između istraživanih grupa u svim istraživanim razdobljima.
Nije uočena statistička značajnost u citotoksičnoj aktivnosti stanica NK između grupa u istraživanim vremenskim točkama.
Koncentracije IL-15, IL-18 i IFN su bile statistički značajno više u bolesnika lječenih on-pump CABG operacijom u usporedbi s bolesnicima liječenih off-pump CABG operacijom 24 i 72 sata nakon operacije u odnosu na prijeoperacijske vrijednosti.
Zaključak
U bolesnika liječenih on-pump CABG operacijom značajno je smanjenudio CD3+ i CD3+CD4+ stanica, izražaj perforina u limfocitima periferne krvi te povećala koncentraciju pro-uplanih citokina (IL-15, IL-18 i IFN u odnosu na bolesnike liječene off-pump CABG operacijom. Ugljikohidratna prijeoperacijska prehrana nije imala statistički značajnog utjecajana promjeneudjela subpopulacijama limfocita i izražaja perforina u istraživanim skupinama bolesnika i istraživanim vremenskim točkama.
Abstract (english) Surgical myocardial revascularization significantly effects changes of immune parameters, especially cell immunity, leading to down-regulation of immune system. Mechanisms of regulation of and down-regulation the immune response and the effect of perioperative feeding on these changes in patients during and early after surgical revascularisation are still not fully understood.
The aim of this study was analyse and compare the influence of peroral feeding on frequency different leukocyte subpopulation, their perforin expression and cytotoxic activity of NK cells as well as cytokine expression (interferon gamma; IFN, interleukin (IL)-15, IL-18) in the blood of patients underwent surgical myocardial revascularisation.
Patients andMethods
In this prospective clinical study 80 patients scheduled for CABG surgery was included in the study. The patients were randomly allocated into four groups (40 in each group): Group 1 underwent on-pump CABG and did not received perioperative nutrition support, patients in Group 2 underwent on-pump CABG and were preoperatively fed, patients in Group 3 underwent off-pump CABG and did not received perioperative nutrition support, while patients in Group 4 underwent off-pump CABG and received preoperative nutrition support. Blood samples (20 ml) were collected immediately before (T1), 24 (T2) and 72 (T3) hours after surgery. Peripheral blood mononuclear cells were isolated by gradient centrifugation and simultaneous labelled by antigens using fluorochrome conjugated monoclonal antibodies. Frequency of T lymphocytes, NK and NKT cells, their subsets as well as their perforin expression was detected and analysed by flow cytometry.
Plama concentrationsofIL-4 and interferon gama (IFNγ) weredetected by ELISA (enzyme linked immunosobrent assay) according to the manufacturer instructions. For NK cells isolation from peripheral blood positive magnetic cell separation method was applied. The cytotoxic activity of NK cells was analysed by flow cytometry.
Results
The frequency of CD3+ and CD3+CD4+,allperforin+ and perforin+CD3+CD8+ cellssignificantly decreased in patients who underwent on-pump CABG when compared to patients who underwent off-pump CABG 24 h after surgery. Frequency of T regulatory cells significantly increased in patients who underwent on-pump CABG when compared to patients who underwent off-pump CABG 24 h after surgery.
The frequency of IL-15+CD3+, IL-15+CD56+ and IL-15+CD3+CD56+ cells significantly increased in patients who underwent off-pump CABG when compared to patients who underwent on-pump CABG 24 h after surgery. The frequency of IFNγ+ cells and cytotoxic activity of NK cells did not differ significantly amnog groups at any time point.
Concentrations of IL-15, IL-18 i IFN significanlty increaed in patients who underwent on-pump CABG surgery when compared to patients who underwent off-pump CABG surgery 24 and 72 h after surgery.
Conclusion
Significntly decreased frequency of CD3+ and CD3+CD4+ cells, perforin expression and increased concentration of proinflammatory cytokines (IL-15, IL-18 i IFNwas found in patients who underwent on-pump CABG surgery when compared to patients who underwent off-pump CABG surgery. Charbohydrate rich perioperative feeding did not significantly effect changes of frequency in different lymphocyte subpopulation and perforin expression among grups at different time points.
Keywords
koronarna bolest srca
perforin
prijeoperacijska peroralna prehrana
stanična imunost.
Keywords (english)
cellular immunity
coronary artery disease
perforin
preopertive peroral nutrition
Language croatian URN:NBN urn:nbn:hr:184:680273 Study programme Title: Biomedicine Postgraduate (doctoral) study programme Type of resource Text File origin Born digital Access conditions Access restricted to students and staff of home institution Terms of use Created on 2019-01-28 11:34:11
