西洛他唑对离体培养大鼠皮层细胞缺血再灌注损伤的保护作用  

Protection of cilostazol on cultured cerebral rats cortical cells models of ischemia-reperfusion injury in vitro

作　　者：王晓坤[1] 梁庆成[2] 吴云[1] 金永华[1] 杨春晓[1] 

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

出　　处：《神经解剖学杂志》2011年第6期653-657,共5页Chinese Journal of Neuroanatomy

基　　金：黑龙江省卫生厅科研课题(2007-333);哈医大二院2007年院博士科研基金项目(BS2007-11)

摘　　要：目的:研究磷酸二酯酶抑制剂西洛他唑对离体培养大鼠皮层细胞缺血再灌注损伤的影响及作用机制。方法:取新生Wistar大鼠皮层细胞进行原代培养,分为5组:正常对照组、西洛他唑组、依达拉奉组、溶剂对照组及缺血再灌注模型组。通过建立糖氧剥离后复糖氧的细胞损伤模型模拟细胞"缺血再灌注损伤",然后进行干预。细胞经4 h糖氧剥离24 h复糖氧培养后,测定培养上清中乳酸脱氢酶(LDH)、丙二醛(MDA)、谷胱甘肽过氧化物酶(GSH-Px)、神经元型一氧化氮合酶(nNOS)、诱导型一氧化氮合酶(iNOS)及一氧化氮(NO)的含量;测定细胞内环磷酸腺苷(cAMP)水平,并通过四唑盐(MTT)比色实验测定细胞活力。结果:西洛他唑和依达拉奉均可减少"缺血再灌注"损伤细胞的LDH和MDA漏出量,提高GSH-Px释放量,降低nNOS、iNOS和NO的水平,升高细胞内cAMP水平,使细胞存活率显著提高;西洛他唑与依达拉奉组相比,LDH、MDA漏出量及GSH-Px的释放量无差别,nNOS、iNOS和NO的水平明显降低,细胞内cAMP水平显著升高,细胞存活率明显提高。结论:西洛他唑对大鼠皮层细胞缺血再灌注损伤具有保护作用,其作用机制可能通过抗氧化、降低nNOS及iNOS的水平,从而降低NO的分泌、升高细胞内cAMP水平来实现的。Objective： To study the effects and possible mechanisms of cilostazol（phosphoiesterase inhibitor） on cultured rats cortical cells models of ischemia-reperfusion injury in vitro.Methods： The cortical cells of the neonatal Wistar rats were cultured in vitro,and then cells were divided randomly into the normal group,cilostazol group,edaravone group,solvent group and ischemia-reperfusion injury（IRI） group.The cell model of ischemia-reperfusion injury was established by regaining oxygen-glucose after oxygen-glucose deprivation mimicking cerebral ischemia-reperfusion injury（IRI）.All the concentrations of lactate dehydrogenase（LDH）,malondialdehyde（MDA）,glutathione peroxidase（GSH-Px）,neuronal nitric oxide synthase（nNOS）,inducible nitric oxide synthase（iNOS） and nitric oxide（NO） in supernatants were measured after 4 h deprivation and 24 h regain of oxygen-glucose.At the same time,the concentrations of cyclic adenosine monophosphate（cAMP） were determined by enzyme linked immunosorbent assay（ELISA） Kit and cells viability was assayed by MTT.Results： Compared with the IRI group,the concentrations of LDH,MDA,nNOS,iNOS and NO were significantly decreased,the levels of GSH-Px and intracellular cAMP were obviously increased,and cells viability were markedly increased in cilostazol group and edaravone group.Compared with the edaravone group,no obvious differences were found in the concentrations of LDH,MDA and GSH-Px,but the concentrations of nNOS,iNOS and NO were significantly decreased in the cilostazol group.The intracellular cAMP levels and cells viability from the cilostazol group were significantly higher than from the edaravone group.Conclusion： The results showed that cilostazol had protective effects on cultured rat cortical neurons and astrocytes in IRI.The mechanisms may be related with resisting oxidation,reducing levels of nNOS and iNOS which results in a reduction of NO secretion,and elevates intracellular cAMP levels in supernatants.

关 键 词：皮层细胞 缺血再灌注损伤 西洛他唑 保护 大鼠 

分 类 号：R741[医药卫生—神经病学与精神病学]