title: Plasma factors in severe early-onset preeclampsia do not substantially alter endothelial gene expression in vitro.
abstract: OBJECTIVE
  Systemic endothelial dysfunction is a central feature in the pathophysiology of preeclampsia. Its cell biologic and molecular basis is poorly understood. One leading hypothesis argues that endothelial dysfunction is caused by (at present largely unknown) circulating factors released from the ischemic placenta. This study investigated the effects of plasma factors of severe, early-onset preeclamptic women versus healthy pregnant women on endothelial gene expression in vitro.
  
  
  METHODS
  Plasma samples were taken from eight severe early-onset preeclamptic women and eight matched pregnant control women. Primary human umbilical vein endothelial cell (HUVEC) and human glomerular microvascular endothelial cell (hGMEC) cultures were incubated with 20% (vol/vol) plasma for 4, 12, and 24 hours. Identical amounts of RNA isolated from HUVEC from three preeclamptic and three control samples were pooled for each time point, and subsequently hybridized on human 60-mer oligonucleotide microarrays containing 17,000 genes. Gene expression levels of vascular cell adhesion molecule-1 (VCAM-1), intercellular adhesion molecule-1 (ICAM-1), interleukin-8 (IL-8), and interleukin-6 (IL-6) in HUVEC and hGMEC were quantified using real-time reverse transcription polymerase chain reaction (RT-PCR).
  
  
  RESULTS
  Microarray analyses of individual genes identified no genes that were up- or down-regulated more than 2.7-fold, and analyses of gene ontologies showed no gene ontology significantly up- or down-regulated in HUVEC by preeclamptic plasma. IL-8 gene expression was modestly induced by preeclamptic plasma after 4, 12, and 24 hours of HUVEC and hGMEC incubation, as identified by real-time RT-PCR. The other genes analyzed did not show altered regulation by preeclamptic plasma factors.
  
  
  CONCLUSIONS
  In vitro, plasma from preeclamptic patients does not substantially alter endothelial gene expression profile. Only modest induction of IL-8 gene expression was observed. These results indicate that mechanisms other than soluble plasma constituents are likely involved in systemic endothelial cell activation in preeclampsia.
authors.affiliation: Department of Obstetrics and Gynecology, Groningen University Institute for Drug Exploration (GUIDE) and Academic Hospital Groningen, Groningen, The Netherlands. r.b.donker@og.azg.nl
authors.forename: Rogier B
authors.initials: RB
authors.lastname: Donker
authors.forename: Sigridur A
authors.initials: SA
authors.lastname: Asgeirsdóttir
authors.forename: Frans
authors.initials: F
authors.lastname: Gerbens
authors.forename: Maria G
authors.initials: MG
authors.lastname: van Pampus
authors.forename: Cees G M
authors.initials: CG
authors.lastname: Kallenberg
authors.forename: Gerard J
authors.initials: GJ
authors.lastname: te Meerman
authors.forename: Jan G
authors.initials: JG
authors.lastname: Aarnoudse
authors.forename: Grietje
authors.initials: G
authors.lastname: Molema
mesh_terms: D000328:Adult; D000941:Antigens / Q000378:metabolism; D002097:C-Reactive Protein / Q000378:metabolism; D004730:Endothelium, Vascular / Q000378:metabolism / Q000502:physiology*; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D020869:Gene Expression Profiling; D006801:Humans; D007231:Infant, Newborn; D018799:Intercellular Adhesion Molecule-1 / Q000096:biosynthesis / Q000097:blood* / Q000235:genetics; D015850:Interleukin-6 / Q000096:biosynthesis / Q000097:blood* / Q000235:genetics; D016209:Interleukin-8 / Q000096:biosynthesis / Q000097:blood* / Q000235:genetics; D020411:Oligonucleotide Array Sequence Analysis; D011225:Pre-Eclampsia / Q000097:blood* / Q000235:genetics; D011247:Pregnancy; D011262:Pregnancy Trimester, Second; D012333:RNA, Messenger / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014471:Umbilical Veins; D019010:Vascular Cell Adhesion Molecule-1 / Q000096:biosynthesis / Q000097:blood* / Q000235:genetics; D014841:von Willebrand Factor / Q000276:immunology
chemical_list: D000941:Antigens; D015850:Interleukin-6; D016209:Interleukin-8; D012333:RNA, Messenger; D019010:Vascular Cell Adhesion Molecule-1; C029417:Von Willebrand antigen; D014841:von Willebrand Factor; D018799:Intercellular Adhesion Molecule-1; D002097:C-Reactive Protein