ABSTRACT
Aim To study the effects of resveratrol(Res)on the release of soluble adhesion molecules from human umbilical vein endothelial cells(HUVECs)induced by N-formyl-methionyl-leucyl-phenylal-anine(fMLP)and the adhesion between neutrophils and HUVECs.Methods The effects of Res on neutrophils adhesion to human umbilical endothelial cell(HUVECs)triggered by fMLP were examined.The soluble intercellular cell adhesive molecule-1(ICAM-1),the soluble vascular cell adhesive molecule-1(VCAM-1)and E-selectin release from fMLP(10 ?mol?L-1)stimulated HUVECs were determined by ELISA kits.The neutrophils isolated from human vein blood were loaded with Fluo-3,a fluorescent indicator,to detect intracellular free calcium concentration([Ca2+]i),and CLA was used as a chemiluminescent indicator to determine superoxide production in neutrophils.Results Res(1~50 ?mol?L-1)significantly inhibited neutrophil adhesion to fMLP-stimulated HUVECs and also obviously downregulated the levels of ICAM-1,VCAM-1 and E-selectin in supernatant of HUVECs culture stimulated by fMLP in the dose-dependent pattern.Res also suppressed fMLP-activated superoxide generation and[Ca2+]i increase in neutrophils.Conclusions Res involved in neutrophil adhesion to HUVECs intermediated by cell adhesive molecules expression trigged by[Ca2+]i and superoxide production in neutrophils,which means a lot to prevent inflammatory diseases.
ABSTRACT
AIM To examine the effect of cilostazol, a no vel selective phosphodiesterase type 3 inhibitor, on adherence between neutrophils and human umbilical e ndothelial cells ( HUVECs ) and investigate its possible mechanisms. MET HODS Confluent HUVECs between 4～6 passages were used and stimulated by l ipopolysaccharide (LPS, 5 mg?L -1 ) with or without coincubation of cilosta zol (1～10 ?mol?L -1 ) for 24 h. Soluble cell adhesion molecules (sCAMs), including vascular cell adhesion molecule-1 (sVCAM-1), intercellular adhesion molecule-1 (sICAM-1) and endothelial leukocyte adhesion molecule-1 (sELAM-1, sE-selectin) in cell culture medium were measured by ELISA. RESULTS Cilostazol (1～10 ?mol?L -1 ) inhibited adherence between neutrophils and HUVECs in a dose- dependent manor. At the same time, cilostazol didn't affect sICAM-1 and sE-sel ectin release from LPS-stimulated HUVECs, but in contrast, it significantly inh ibited sVCAM-1 production under the same experiment condition, and this effect was canceled by H-89, an inhibitor of protein kinase A ( PKA ). CONCLUS ION Cilostazol significantly inhibits adherence between neutrophils and H UVECs, and downregulates sVCAM-1 release from LPS-activated HUVECs, and these effects on cytokine-challenged endothelial cells might be via a PKA-dependent pathway. The present result suggests that cilostazol partially eliminates some o f the adherent reactions of HUVECs to LPS, a deleterious cytokine, and it is rea sonable to consider that cilostazol might be a strategy for preventing atheroscl erosis and other cardiovascular diseases.