Expression of inflammation-related genes in endothelial cells is not directly affected by microparticles from preeclamptic patients

J Lab Clin Med. 2006 Jun;147(6):310-20. doi: 10.1016/j.lab.2006.02.004.

Abstract

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.

Aim: 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.

Conclusion: 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.

MeSH terms

  • Adult
  • Chemokine CCL2 / genetics
  • Chemokine CCL2 / immunology
  • Cytokines / genetics*
  • Cytokines / immunology*
  • Endothelial Cells / immunology*
  • Female
  • Gene Expression Regulation / immunology
  • Humans
  • In Vitro Techniques
  • Interleukin-1 / genetics
  • Interleukin-1 / immunology
  • Interleukin-2 / genetics
  • Interleukin-2 / immunology
  • Interleukin-6 / genetics
  • Interleukin-6 / immunology
  • Interleukin-8 / genetics
  • Interleukin-8 / immunology
  • NF-kappa B / genetics
  • NF-kappa B / immunology
  • Particle Size
  • Pre-Eclampsia / immunology*
  • Pre-Eclampsia / physiopathology*
  • Pregnancy
  • RNA, Messenger / analysis
  • Reverse Transcriptase Polymerase Chain Reaction
  • Umbilical Veins / cytology

Substances

  • Chemokine CCL2
  • Cytokines
  • Interleukin-1
  • Interleukin-2
  • Interleukin-6
  • Interleukin-8
  • NF-kappa B
  • RNA, Messenger