Related Experiment Video
Updated: Aug 8, 2026
Screening Assays to Characterize Novel Endothelial Regulators Involved in the Inflammatory Response
Published on: September 15, 2017
Expression of inflammation-related genes in endothelial cells is not directly affected by microparticles from
Christianne Anne Rachel Lok1, Anita N Böing, Pieter H Reitsma
1Department of Obstetrics, Academic Medical Center, Amsterdam, The Netherlands. c.a.lok@amc.uva.nl
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.
Insights
Microparticles (MPs) from preeclamptic patients did not directly alter inflammation-related gene expression in endothelial cells. This suggests MPs may not cause endothelial dysfunction in preeclampsia through this specific mechanism.
Area of Science:
- Obstetrics and Gynecology
- Cardiovascular Biology
- Immunology
Background:
- Preeclampsia is characterized by inflammation and endothelial dysfunction.
- Microparticles (MPs) are implicated in linking inflammation and endothelial dysfunction.
- MPs can stimulate pro-inflammatory cytokine production and impair endothelial function.
Purpose of the Study:
- To investigate how MPs from preeclamptic patients affect inflammation-related gene expression in human endothelial cells.
- To determine if MPs directly influence the inflammatory profile of endothelial cells.
Main Methods:
- Human umbilical vein endothelial cells (HUVECs) were exposed to MP fractions from preeclamptic, normotensive pregnant, and non-pregnant individuals.
- Multiplex ligation-dependent probe amplification (MLPA) and real-time PCR were used to analyze RNA expression.
- Interleukin (IL)-1alpha served as a positive control for inflammatory gene induction.
Main Results:
- IL-1alpha significantly upregulated several inflammation-related genes, including IL-1alpha, IL-2, IL-6, IL-8, NF-kappaB family members, and monocyte chemotactic protein-1.
- Tissue factor expression was transiently increased by IL-1alpha.
- MP fractions from all groups did not alter the expression of inflammation-related genes or adhesion receptors in HUVECs.
Conclusions:
- Multiplex ligation-dependent probe amplification (MLPA) is a validated method for assessing endothelial cell inflammatory status.
- The study suggests that MPs from preeclamptic patients are unlikely to induce endothelial dysfunction by directly modulating inflammation-related gene expression.
- Further research is needed to elucidate the precise mechanisms by which MPs contribute to preeclampsia pathophysiology.
