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.
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: J X Chen; L C Berry; B W Christman; M Tanner; P R Myers; B O Meyrick Journal: Am J Physiol Lung Cell Mol Physiol Date: 2001-03 Impact factor: 5.464