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

中国临床药理学与治疗学 ›› 2003, Vol. 8 ›› Issue (6): 621-626.

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

罗景慧^1,2, 林永成², 陈志良¹, 渡边裕司³, 尾门关真理子⁴, 林秀晴⁴

¹南方医院药学部,广州 510515,广东;
²中山大学化学系,广州 510275,广东;
³浜松医科大学临床药理学研究室,⁴浜松医科大学第三内科,浜松 431-3192,日本

收稿日期:2003-05-24 接受日期:2003-08-14 出版日期:2003-12-26 发布日期:2020-11-19

Effects of aPhosphodiesterase 3 inhibitor, cilostazol on soluble adhesion molecules release from TNF-α-stimulated human umbilical endothelial cells

LUO Jing-Hui^1,2, LIN Yong-Cheng², CHEN Zhi-LIang¹, WATANABE Hiroshi³, OZEKI Mariko⁴, HAYASHI Hideharu⁴

¹Pharmaceutical Department,Nanfang Hospital,Guangzhou 510515,Guangdong,China;
²Department of Chemistry,Zhongshan University,Guangzhou 510275,Guangdong,China;
³Department of ClinicalPharmacology and Therapeutics and⁴Internal Medicine Ⅲ,Hamamatsu University School of Medicine,Hamamatsu 431-3192,Japan

Received:2003-05-24 Accepted:2003-08-14 Online:2003-12-26 Published:2020-11-19
Contact: LUO Jing-Hui,female,Ph.D,engaged in pharmacology and marine natural productschemistry.Tel:020-61640322 　Fax:020-61648236 　E-mail:yangyingluohui@hotmail.com

摘要/Abstract

摘要： 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(10μg·L^-1)刺激细胞,并与西洛他唑(1 ~ 10μmol·L^-1)共培养24 h,取培养上清,通过ELISA 法测定可溶性血管细胞粘附分子-1(sVCAM-、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 ~ 10μmol·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 24 h,低浓度(1μmol·L^-1)可显著改善细胞生长状态;高浓度(30μmol·L^-1)表现为抑制,而中浓度(10μmol·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,因此在动脉粥样硬化以及其它心血管疾病的治疗中具有潜在的应用价值。

关键词: 磷酸二酯酶3 型抑制剂, 西洛他唑, 可溶性粘附分子, 人脐静脉内皮细胞

Abstract: AIM: To examine the effects of cilostazol,a novel selectivePhosphodiesterase type 3 inhibitor,on soluble cell adhesion molecules(sCAMs)released from tumor necrosis factor-α(TNF-α)-stimulated human umbilical endothelial cells(HUVECs),and to investigate thePossible mechanisms of these effects of cilostazol.METHODS: Confluent HUVECs between 4-6Passages were used and stimulated by TNF-α(10μg·L^-1)with or without coincubation of cilostazol(1 -10μmol·L^-1)for 24 h.Soluble vascular cell adhesion molecule-1(sVCAM-1),soluble intercellular adhesion molecule-1(sICAM-1)and soluble endothelialleukocyte adhesion molecule-1(sELAM-1,sE-selectin)in cell culture medium were measured by ELISA,and availability of cells was detected by MTT assay.RESULTS: Cilostazol(1 -10μmol·L^-1)did not affect sICAM-1 and sE-selectin released from HUVECs,but in contrast,it significantly inhibited theProduction of TNF-α-induced sVCAM-1,and this effect was canceled bylω-nitro-L-arginine methyl ester(l^ω-NAME,0.1μmol·L^-1),a nonselective nitro oxide synthase(NOS)inhibitor.MTT assay indicated that the treatment of HUVECs with cilostazol(1 -30μmol·L^-1)for 24 h affected cell availability in a complexPattern.It was increased at alow dose of cilostazol(1μmol·L^-1),but it was decreased at a relatively high dosage,30μmol·L-1.And at the medium dosage of cilostazol,10μmol·L^-1,cell availability was almost unaffected.CONCLUSIONS: Cilostazol significantly inhibits sVCAM-1 released from TNF-α-activated HUVECs,and the effect on cytokine-challenged endothelial cells might have some relationshiPwith activating NOS,andPossibly,it is via a NO-dependentPathway.ThePresent result suggests that cilostazolPartially eliminates some of the adherent reactions of HUVECs to TNF-α,a deleterious cytokine,and to some extent,it might have thePotential toPrevent atherosclerosis and other cardiovascular diseases.

Key words: phosphodiesterase 3 inhibitor, cilostazol, soluble cell adhesion molecules, human umbilical endothelial cells

中图分类号:

R965

罗景慧, 林永成, 陈志良, 渡边裕司, 尾门关真理子, 林秀晴. 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[J]. 中国临床药理学与治疗学, 2003, 8(6): 621-626.

LUO Jing-Hui, LIN Yong-Cheng, CHEN Zhi-LIang, WATANABE Hiroshi, OZEKI Mariko, HAYASHI Hideharu. Effects of aPhosphodiesterase 3 inhibitor, cilostazol on soluble adhesion molecules release from TNF-α-stimulated human umbilical endothelial cells[J]. Chinese Journal of Clinical Pharmacology and Therapeutics, 2003, 8(6): 621-626.

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