Abstract

A rapid, cost-effective and sensitive method of Northern analysis using specific oligonucleotide probes combined with chemiluminescence was used to detect adhesion molecule transcripts with as little as 50 ng of total RNA from activated primary human umbilical vein endothelial cells (HUVEC). Activation of cultured HUVEC with the inflammatory cytokine interleukin 1β (IL-1β) resulted in distinct time- and dose-dependent patterns of expression of ICAM-1, E-Selectin and VCAM-1 transcripts. The levels of all three transcripts were maximal after 2.5 hours of stimulation with IL-1β. However, the upregulation of ICAM-1 mRNA levels in response to IL-1β was much more stable than the transient induction of E-selectin and VCAM-1 transcripts. In addition, we have found that eicosapentaenoic acid (EPA) and docosapentaenoic acid (DHA) ω3 fatty acids that have been reported to modulate the adhesive properties of cells involved in immune/inflammatory responses do not affect the expression of ICAM-1, VCAM-1 or E-selectin in resting HUVEC. However, EPA (65 μM) or DHA (65 μM) do attenuate the induction of each of these adhesion molecules in IL-1β-activated HUVEC.

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