西洛他唑对大鼠脑微血管内皮细胞缺血再灌注损伤的保护作用  

The protective roles of cilostazol on cerebral microvascular endothelial cells in ischemia reperfusion model in vitro

作　　者：吴云[1] 梁庆成[2] 王晓坤[1] 丛林[1] 

机构地区：[1]哈尔滨医科大学附属第二医院神经内科,150086 [2]华中科技大学同济医学院在读博士,430030

出　　处：《中国神经免疫学和神经病学杂志》2011年第2期106-109,共4页Chinese Journal of Neuroimmunology and Neurology

基　　金：2008年黑龙江省卫生厅科研课题基金资助项目(No.2007-333);哈尔滨医科大学附属第二医院2007年院博士科研基金资助项目(No.BS2007-11)

摘　　要：目的研究西洛他唑对大鼠脑微血管内皮细胞缺血再灌注损伤的保护作用机制。方法以原代方法培养大鼠脑微血管内皮细胞并传代,将第3代传代细胞随机分为正常对照组、西洛他唑组、溶剂对照组和缺血再灌注模型组(再灌模型组)4组,对后3组大鼠建立脑微血管内皮细胞糖氧剥夺后复糖氧模型,模拟"缺血再灌注"过程,并对西洛他唑组和溶剂对照组在造模同时分别以终浓度1×10-5mmol/L西洛他唑〔二甲基亚砜(DMSO)为溶剂〕和0.05%(体积分数)DMSO进行干预。糖氧剥夺3 h复糖氧24 h后,测定各组细胞上清液中诱导型一氧化氮合酶(iNOS)和内皮型一氧化氮合酶(eNOS)水平及细胞内环磷腺苷酸(cAMP)水平,并以四唑盐(MTT)比色实验测定细胞活力。结果西洛他唑组与再灌模型组比较,eNOS水平明显提高,iNOS水平明显降低,cAMP水平及细胞存活率显著提高(均P<0.05)。结论西洛他唑对大鼠脑微血管内皮细胞的缺血再灌注损伤有保护作用,其作用机制可能与促进eNOS水平升高和iNOS水平降低有关。Objective To study the protective mechanisms of cilostazol on cerebral microvascular endothelial cells in ischemia reperfusion models in vitro.Methods Cerebral microvascular endothelial cells of the Wistar rats were cultured and subcultured in vitro.All the 3rd generation cells were divided randomly into the normal group,the cilostazol treatment group,the solvent group and the ischemia-reperfusion injury（IRI） group.The cell model of ischemia reperfusion was established by regaining oxygen-glucose after oxygen-glucose deprivation mimicking cerebral IRI.At the same time,the cilostazol group was treated with cilostazol（1×10-5mmol/L） in dimethyl sulfoxide（DMSO,0.05%） and the solvent group was treated with 0.05% DMSO,only.Three hours after oxygen-glucose deprivation and 24 h after regain of oxygen-glucose,the levels of endothelial nitric oxide synthase（eNOS） and inducible nitric oxide synthase（iNOS） in supernatants were measured.At the same time,the concentrations of cyclic adenosine monophosphate（cAMP） were determined by enzyme linked immunosorbent assay（ELISA）,and cell viability was assayed by MTT.Results Compared with the IRI group,the level of iNOS significantly decreased（P0.05）,the levels of eNOS and intracellular cAMP obviously increased（P0.05）,and cell viability was marked increased in the cilostazol group（P0.05）.Conclusions The results showed that cilostazol had protective effects on cultured rat cerebral microvascular endothelial cells in ischemia-reperfusion injury models.The mechanisms may be that cilostzaol could increase the levels of eNOS and reduce the levels of iNOS.

关 键 词：西洛他唑 脑微血管内皮细胞 缺血再灌注损伤 保护 

分 类 号：R743.3[医药卫生—神经病学与精神病学]