Urocortin induced expression of COX-2 and ICAM-1 via corticotrophin-releasing factor type 2 receptor in rat aortic endothelial cells.

Abstract

BACKGROUND AND PURPOSE:

Our previous study showed that urocortin (Ucn1) exacerbates the hypercoagulable state and vasculitis in a rat model of sodium laurate-induced thromboangiitis obliterans. Furthermore, the inflammatory molecules COX-2 and ICAM-1 may participate in this effect. In the present study, the effects of Ucn1 on COX-2 and ICAM-1 expression in lipopolysaccharide (LPS)-induced rat aortic endothelial cells (RAECs) were investigated and the mechanisms involved explored.

EXPERIMENTAL APPROACH:

RAECs were isolated from adult male Wistar rats, and identified at the first passage. Experiments were performed on cells, from primary culture, at passages 5-8. The expression of COX-2 and ICAM-1 at both mRNA and protein levels was determined by semi-quantitative RT-PCR and Western blot analysis. Levels of PGE(2) and soluble ICAM-1 (sICAM-1) in culture medium were measured by enzyme-linked immunosorbent assay. Furthermore, the phosphorylation status of p38MAPK, ERK1/2, JNK, Akt and NF-kappaB was analysed by Western blot; nuclear translocation of NF-kappaB was observed by immunofluorescence.

KEY RESULTS:

Ucn1 augmented LPS-induced expression of COX-2 and ICAM-1 in RAECs in a time- and concentration-dependent manner. Ucn1 increased PGE(2) and sICAM-1 levels. These effects were abolished by the CRF(2) receptor antagonist, antisauvagine-30, but not by the CRF(1) receptor antagonist, NBI-27914. Moreover, Ucn2 activated p38MAPK and augmented NF-kappaB nuclear translocation and phosphorylation, whereas ERK1/2, JNK and Akt pathways were not involved in this process.

CONCLUSIONS AND IMPLICATIONS:

These findings suggest that Ucn1 exerts pro-inflammatory effects by augmenting LPS-induced expression of COX-2 and ICAM-1 in RAECs via CRF(2) receptors and the activation of p38MAPK and NF-kappaB.

• Authors: Rongjian Zhang, Youhua Xu, Hong Fu, Juejin Wang, Lai Jin, Shengnan Li
• Journal: British journal of pharmacology (Br J Pharmacol) Vol. 158 Issue 3 Pg. 819-29 (Oct 2009) ISSN: 1476-5381 [Electronic] England
• PMID: 19694731 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)

Chemical References

• 2-methyl-4-(N-propyl-N-cycloproanemethylamino)-5-chloro-6-(2,4,6-trichloranilino)pyrimidine
• Aniline Compounds
• CRF receptor type 2
• Lipopolysaccharides
• NF-kappa B
• Peptide Fragments
• Pyrimidines
• Receptors, Corticotropin-Releasing Hormone
• Urocortins
• antisauvagine 30
• Intercellular Adhesion Molecule-1
• CRF receptor type 1
• Cyclooxygenase 2
• Ptgs2 protein, rat
• p38 Mitogen-Activated Protein Kinases
• Dinoprostone

Topics

• Active Transport, Cell Nucleus
• Aniline Compounds (pharmacology)
• Animals
• Aorta (cytology, metabolism)
• Cyclooxygenase 2 (biosynthesis)
• Dinoprostone (metabolism)
• Endothelial Cells (metabolism)
• Enzyme Activation
• Intercellular Adhesion Molecule-1 (biosynthesis)
• Lipopolysaccharides (pharmacology)
• Male
• NF-kappa B (metabolism)
• Peptide Fragments (pharmacology)
• Phosphorylation
• Pyrimidines (pharmacology)
• Rats
• Rats, Wistar
• Receptors, Corticotropin-Releasing Hormone (antagonists & inhibitors)
• Urocortins (physiology)
• p38 Mitogen-Activated Protein Kinases (metabolism)