西洛他唑对磷脂多糖诱导的粘附及粘附分子释放的影响

目的　研究磷酸二酯酶 3型抑制剂西洛他唑对磷脂多糖 (LPS)诱导的粘附及血管内皮细胞 (ECs)释放可溶性细胞粘附分子 (sCAMs)的影响。方法　体外培养第 4～ 6代人脐静脉血管内皮细胞 (HUVECs) ,以LPS(5mg·L- 1)刺激 ,并与西洛他唑 (1～ 10 μmol·L- 1)共培养 2 4h ,观察西洛他唑对由LPS诱导的HUVECs与中性粒细胞之间的粘附的影响 ;另取培养上清 ,以ELISA法测定HUVECs释放可溶性血管细胞粘附分子 1(sVCAM 1)、细胞间粘附分子 1(sICAM 1)以及E 选择素 (sE selectin ,sELAM 1)。结果　西洛他唑抑制由LPS诱导的HUVECs与中性粒细胞之间的粘附以及HUVECs释放sVCAM 1,而对sICAM 1和sE 选择素无影响。并且 ,该药对于sVCAM 1的抑制作用被蛋白激酶A(PKA)抑制剂H 89所阻断。结论　西洛他唑对由细胞因子LPS诱导的粘附反应以及ECs释放sVCAM 1有抑制作用 ,后者可能与PKA依赖性通路相关

Effects of cilostazol on LPS-stimulated adhesion and soluble adherent molecules release

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 μmol·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 μmol·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.