Naslov (srp)

Markeri sinteze i apsorpcije holesterola i sfingolipidni profil u preeklampsiji : doktorska disertacija

Autor

Antonić Krejović, Tamara, 1993-

Doprinosi

Ardalić, Daniela, 1967-
Miković, Željko, 1960-
Stefanović, Aleksandra, 1977-
Zeljković, Aleksandra, 1974-

Opis (srp)

Uprkos napretku u razjašnjavanju patofizioloških mehanizama uključenih u razvoj preeklampsije,jedina efikasna terapija je porođaj, a rizik za razvoj preeklampsije se najčešće procjenjuje na osnovudemografskih karakteristika i faktora rizika prisutnih kod majke, koji imaju niske pozitivne prediktivnevrijednosti. Kako kardiovaskularne bolesti i preeklampsija dijele niz zajedničkih faktora rizika, aporemećaji u metabolizmu lipida i lipoproteina se smatraju ključnim faktorima u patogeneziateroskleroze i razvoju kardiovaskularnih bolesti, nameće se potreba za detaljnijim ispitivanjempromjena u metabolizmu lipida i njihovog značaja u predikciji rizika za razvoj preeklampsije. Cilj ovogistraživanja bio je longitudinalno praćenje specifičnih komponenti lipidoma kod žena savisokorizičnom trudnoćom, analiza razlika u specifičnim elementima lipidoma između trudnica sa i bezpreeklampsije, i procjena dijagnostičke tačnosti ispitivanih parametara.U studiju je uključeno 90 trudnica sa visokim rizikom sa razvoj preeklampsije, a koje su se javile radipraćenja trudnoće u Ginekološko-akušersku kliniku „Narodni Front“ u Beogradu. Na osnovuprimarnog ishoda trudnoće, ispitanice su klasifikovane u grupu trudnica sa visokorizičnom trudnoćom(RG), koja nije razvila preeklampsiju, i grupu trudnica sa preeklampsijom (PG). U prvoj faziistraživanja određeni su osnovni klinički i biohemijski parametri i osnovni parametri lipidnog profilatokom prvog, drugog, trećeg trimestra i pred porođaj. Koncentracije neholesterolskih sterola (NHS) userumu i lipoproteinskim česticama visoke gustine (HDL) su određene metodama tečne hromatografijesa tandem masenom detekcijom (HPLC-MS/MS), prethodno implementiranim i validiranim ulaboratoriji Katedre za medicinsku biohemiju, a izabrani sfingolipidi HPLC-MS/MS metodomrazvijenom za potrebe ove studije, takođe, u četiri tačke tokom gestacije. Masena koncentracijaapolipoproteina M (apoM) je određivana tehnikom enzimskog imunoeseja. Aktivnosti lecitin-holesterolaciltransferaze (LCAT) i holesterol-estar transfernog proteina (CETP) su procijenjene na osnovu brzinestvaranja i prenosa holesterol estara. Aktivnost paraoksonaze 1 (PON1) mjerena je kinetičkikorišćenjem paraoksona kao supstrata.U okviru studije uspješno je optimizovana i validirana HPLC-MS/MS metoda za kvantifikacijusfingozina, sfinganina, sfingozin-1-fosfata (S1P), sfinganin-1-fosfata, ceramida C16:0 (Cer C16:0) iC24:0 (Cer C24:0), i sfingomijelina C16:0 (SM C16:0). Zbog izraženog efekta matriksa, prilikomkonstrukcije kalibracionih krivih kao matriks je korišten pool humane plazme. Varijacije unutar serijasu bile 3,8-18,8%, a između serija 4,1-19,4%. Tačnost se kretala u opsegu od ± 15% od nominalnevrijednosti. Koncentracije triglicerida su bile značajno više u PG u odnosu na RG (p <0,05 u svimispitivanim tačkama), dok je koncentracija HDL holesterola bila značajno niža u PG u 2. trimestru upoređenju sa RG (p <0,05). Povećanje koncentracije holesterola u RG je bilo praćeno značajnimpovećanjem koncentracija holesterolskih prekursora dezmosterola (p <0,001), 7-dehidroholesterola (p<0,05) i latosterola (p <0,001). S druge strane, u PG uprkos povećanju koncentracije ukupnogholesterola, nisu uočene značajne promjene u koncentracijama dezmosterola (p = 0,427) i 7-dehidroholesterola (p =0,368), već samo u koncentraciji latosterola (p <0,05). Porast koncentracijeholesterolskh prekursora krajem trudnoće u RG bio je praćen značajnim padom koncentracija markeraapsorpcije kampesterola (p <0,05) i β-sitosterola (p <0,05), dok u PG uprkos porastu koncentracijelatosterola nije bilo značajnih promjena u koncentracijama biljnih sterola (p =0,364 za kampesterol i p=0,426 za β-sitosterol), što upućuje na djelimično izmjenjenu homeostazu holesterola u PG. Uočeni surazličiti obrasci u sfingolipidnim profilima između ispitivanih grupa. Koncentracije S1P u RG su sepovećavale od 1. trimestra do porođaja (p <0,001)...

Opis (srp)

Medicinske nauke – Farmacija - Medicinska biohemija / Medical Sciences – Pharmacy- Medical Biochemistry Datum odbrane: 13.06.2025.

Opis (eng)

Despite advancements in elucidating the pathophysiological mechanisms involved in the developmentof preeclampsia, delivery remains the only effective therapy. The risk of developing preeclampsia ismost commonly assessed based on demographic characteristics and maternal risk factors, which havelow positive predictive values. Since cardiovascular diseases and preeclampsia share numerous riskfactors, and lipid and lipoprotein metabolism disorders are considered key factors in the pathogenesisof atherosclerosis and the development of cardiovascular diseases, there is a need for a more detailedinvestigation of lipid metabolism changes and their significance in predicting the risk of preeclampsia.The aim of this study was to longitudinally monitor specific components of the lipidome in womenwith high-risk pregnancies, analyze differences in specific lipidome elements between women with andwithout preeclampsia, and evaluate the diagnostic accuracy of the examined parameters.The study included 90 pregnant women at high risk for developing preeclampsia, who were monitoredat the Gynecology and Obstetrics Clinic "Narodni Front" in Belgrade. Based on the primary pregnancyoutcome, participants were classified into a high-risk pregnancy group (RG) that did not developpreeclampsia and a preeclampsia group (PG). In the first phase of the study, basic clinical andbiochemical parameters, as well as lipid profile parameters, were determined during the first, second,and third trimesters, and before delivery. Concentrations of non-cholesterol sterols (NCS) in serum andhigh-density lipoprotein particles (HDL) fraction were measured using liquid chromatography-tandemmass spectrometry (HPLC-MS/MS) methods previously implemented and validated in the laboratoryof the Department of Medical Biochemistry. Selected sphingolipids were measured using an HPLCMS/MS method developed specifically for this study, also at four gestational points. The massconcentration of apolipoprotein M (apoM) was determined using an enzyme immunoassay technique.The activities of lecithin-cholesterol acyltransferase (LCAT) and cholesterol ester transfer protein(CETP) were assessed based on the rates of cholesterol ester formation and transfer. Paraoxonase 1(PON1) activity was measured kinetically using paraoxon as a substrate.In this study, an HPLC-MS/MS method was successfully optimized and validated for quantifyingsphingosine, sphinganine, sphingosine-1-phosphate (S1P), sphinganine-1-phosphate, ceramide C16:0(Cer C16:0), ceramide C24:0 (Cer C24:0), and sphingomyelin C16:0 (SM C16:0). Due to a pronouncedmatrix effect, a pool of human plasma was used as the matrix for constructing calibration curves. Intraassayvariations ranged from 3.8% to 18.8%, while inter-assay variations were between 4.1% and19.4%, with accuracy within ±15% of the nominal values. Triglyceride concentrations weresignificantly higher in the PG compared to the RG (p < 0.05 at all points), while HDL cholesterollevels were significantly lower in PG during the 2nd trimester compared to RG (p < 0.05). In RG, anincrease in cholesterol concentrations was accompanied by significant increases in cholesterolprecursors desmosterol (p < 0.001), 7-dehydrocholesterol (p < 0.05), and lathosterol (p < 0.001). In PG,despite rising total cholesterol levels, no significant changes were observed for desmosterol (p = 0.427)or 7-dehydrocholesterol (p = 0.368), with only lathosterol showing a significant increase (p < 0.05).Late pregnancy increases in cholesterol precursors in RG were associated with significant decreases inabsorption markers campesterol (p < 0.05) and β-sitosterol (p < 0.05). In contrast, PG showed nosignificant changes in phytosterols’ concentrations despite lathosterol increases (p = 0.364 forcampesterol and p = 0.426 for β-sitosterol), indicating altered cholesterol homeostasis in PG. Distinctsphingolipid profiles were observed between the groups. S1P concentrations in RG increased from the1st trimester to delivery (p < 0.001)...

Jezik

srpski

Datum

2025

Licenca

Ovo delo je licencirano pod uslovima licence
Creative Commons CC BY-NC-SA 3.0 AT - Creative Commons Autorstvo - Nekomercijalno - Deliti pod istim uslovima 3.0 Austria License.

http://creativecommons.org/licenses/by-nc-sa/3.0/at/legalcode

Predmet

OSNO - Opšta sistematizacija naučnih oblasti, Farmacija

preeklampsija, lipidni status, neholesterolski steroli, prekursori holesterola, fitosteroli, HDL, sfingolipidi, sfingozin-1-fosfat, ceramidi, sfingomijelin

615:618.3(043.3)

OSNO - Opšta sistematizacija naučnih oblasti, Farmacija

preeclampsia, lipid profile, non-cholesterol sterols, cholesterol precursors, phytosterols, HDL, sphingolipids, sphingosine-1-phosphate, ceramides, sphingomyelin