Expression of inflammation-related genes in endothelial cells is not directly affected by microparticles from preeclamptic patients.
BACKGROUND
Inflammation and endothelial dysfunction are prominent in preeclampsia. Microparticles (MPs) may link these processes, as MPs induce the production of pro-inflammatory cytokines by endothelial cells and cause endothelial dysfunction.


OBJECTIVE
To study changes in expression of inflammation-related genes in human endothelial cells in response to MPs from preeclamptic patients.


METHODS
Human umbilical vein endothelial cells (HUVECs) were incubated for various time intervals in the absence or presence of isolated MP fractions from preeclamptic patients (n = 3), normotensive pregnant women (n = 3), non-pregnant controls (n = 3), and interleukin (IL)-1alpha as a positive control. Total RNA was isolated and used for multiplex ligation-dependent probe amplification (MLPA) and real-time polymerase chain reaction (PCR).


RESULTS
IL-1alpha enhanced the expression of IL-1alpha, IL-2, IL-6, and IL-8; nuclear factor of kappa light chain enhancer in B-cells (NFkappaB)-1, NFkappaB-2, and NFkappaB-inhibitor; cyclin-dependent kinase inhibitor and monocyte chemotactic protein-1; and transiently increased tissue factor expression. RNA expression of inflammation-related genes and genes encoding adhesion receptors, however, were unaffected by any of the MP fractions tested.


CONCLUSIONS
MLPA is a suitable assay to test the inflammatory status of endothelial cells, because incubation with IL-1alpha triggered substantial changes in RNA expression in endothelial cells. Taken together, it seems unlikely that MPs from preeclamptic patients induce endothelial dysfunction by directly affecting the expression of inflammation-related genes in these cells.
Lok
Christianne Anne Rachel
CA
Department of Obstetrics, Academic Medical Center, Amsterdam, The Netherlands. c.a.lok@amc.uva.nl
Böing
Anita N
AN
Reitsma
Pieter H
PH
van der Post
Joris A M
JA
van Bavel
Ed
E
Boer
Kees
K
Sturk
Augueste
A
Nieuwland
Rienk
R
D000328:Adult; D018932:Chemokine CCL2 / Q000235:genetics / Q000276:immunology; D016207:Cytokines / Q000235:genetics* / Q000276:immunology*; D042783:Endothelial Cells / Q000276:immunology*; D005260:Female; D005786:Gene Expression Regulation / Q000276:immunology; D006801:Humans; D066298:In Vitro Techniques; D007375:Interleukin-1 / Q000235:genetics / Q000276:immunology; D007376:Interleukin-2 / Q000235:genetics / Q000276:immunology; D015850:Interleukin-6 / Q000235:genetics / Q000276:immunology; D016209:Interleukin-8 / Q000235:genetics / Q000276:immunology; D016328:NF-kappa B / Q000235:genetics / Q000276:immunology; D010316:Particle Size; D011225:Pre-Eclampsia / Q000276:immunology* / Q000503:physiopathology*; D011247:Pregnancy; D012333:RNA, Messenger / Q000032:analysis; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014471:Umbilical Veins / Q000166:cytology
D018932:Chemokine CCL2; D016207:Cytokines; D007375:Interleukin-1; D007376:Interleukin-2; D015850:Interleukin-6; D016209:Interleukin-8; D016328:NF-kappa B; D012333:RNA, Messenger