磷酸二酯酶 3型抑制剂西洛他唑对TNF-α诱导人脐静脉内皮细胞释放可溶性粘附分子的影响(英文)

Effects of a phosphodiesterase 3 inhibitor, ci lostazol on soluble adhesion molecules release from TNF-α-stimulated huma n umbilical endothelial cells

目的 :研究新型磷酸二酯酶 3型抑制剂西洛他唑 (cilostazol)对TNF α诱导人脐静脉内皮细胞(HUVECs)释放可溶性细胞粘附分子 (sCAMs)的影响 ,并探讨其作用机制。方法 :体外培养第 4～ 6代HUVECs,以TNF α (10 μg·L-1)刺激细胞 ,并与西洛他唑 (1～ 10 μmol·L-1)共培养 2 4h ,取培养上清 ,通过ELISA法测定可溶性血管细胞粘附分子 1(sV CAM 1)、细胞间粘附分子 1(sICAM 1)以及E 选择素(sELAM 1,sE selectin) ,并以四唑蓝 (MTT)法考察细胞生长状态。结果 :1～ 10 μmol·L-1的西洛他唑对TNF α诱导的HUVECs释放sICAM 1和sE selectin无明显影响 ,但显著抑制sVCAM 1的生成 ,并且该作用被一种非选择性一氧化氮合成酶 (NOS)抑制剂Lω NAME(0 .1μmol·L-1)所阻断。MTT法测定结果显示 ,西洛他唑作用于HUVECs 2 4h ,低浓度(1μmol·L-1)可显著改善细胞生长状态 ;高浓度(30 μmol·L-1)表现为抑制 ,而中浓度 (10 μmol·L-1)对细胞生长状态几乎无影响。结论 :西洛他唑显著抑制由TNF α诱导的HUVECs释放sVCAM 1,该作用可能与激活NOS并通过NO依赖性通路介导有关 ,提示该药可在一定程度上对抗由细胞因子所引起的部分粘附反应 。

AIM: To examine the effects of cilostazol, a novel s el ective phosphodiesterase type 3 inhibitor, on soluble cell adhesion molecules (s CAMs) released from tumor necrosis factor-α (TNF-α)-stimulated human umbili cal endothelial cells (HUVECs), and to investigate the possible mechanisms of th ese effects of cilostazol. METHODS: Confluent HUVECs between 4- 6 passages were used and stimulated by TNF-α ( 10 μg·L -1 ) with or w ithout coincubation of cilostazol (1- 10 μmol·L -1 ) for 24 h. Soluble vascular cell adhesion molecule-1 (sVCAM-1), soluble intercellular adhesion mo lecule-1 (sICAM-1) and soluble endothelial leukocyte adhesion molecule-1 (sEL AM-1, sE-selectin) in cell culture medium were measured by ELISA, and availabi lity of cells was detected by MTT assay. RESULTS: Cilostazol (1- 10 μmol·L -1 ) did not affect sICAM-1 and sE-selectin released from HUVECs, but in contrast, it significantly inhibited the production of TNF-α-i nduced sVCAM-1, and this effect was canceled by L ω-nitro- L-arginine methyl ester (L ω-NAME, 0.1 μmol·L -1 ), a nonselective nitro oxide synthase (NOS) inhibitor. MTT assay indicated that the treatment of HUVECs with cilostazol (1- 30 μmol·L -1 ) for 24 h affected cell availability in a complex pattern. It was increased at a low dose of cilostazol ( 1 μmol· L -1 ), but it was decreased at a relatively high dosage, 30 μmol·L -1 . And at the medium dosage of cilostazol, 10 μmol·L -1 , cell availability was almost unaffected. CONCLUSIONS: Cilostazol sign ificantly inhibits sVCAM-1 released from TNF-α-activated HUVECs, and the eff ect on cytokine-challenged endothelial cells might have some relationship with activating NOS, and possibly, it is via a NO-dependent pathway. The present res ult suggests that cilostazol partially eliminates some of the adherent reactions of HUVECs to TNF-α, a deleterious cytokine, and to some extent, it might have the potential to prevent atherosclerosis and other cardiovascular diseases.