西洛他唑对大鼠皮层细胞糖氧剥离损伤的保护作用  

Protection of cilostazol on cultured cerebral cortical cells deprived of oxygen and glucose in rats

作　　者：吴云[1] 王晓坤[1] 梁庆成[2] 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,150086 [2]华中科技大学同济医学院

出　　处：《卒中与神经疾病》2010年第5期259-262,266,共5页Stroke and Nervous Diseases

基　　金：2008年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)共同资助

摘　　要：目的探讨磷酸二酯酶抑制剂西洛他唑对糖氧剥离后大鼠皮层细胞培养的影响及作用机制。方法原代混合培养大鼠皮层细胞,建立糖氧剥离的细胞损伤模型模拟细胞"缺血损伤",然后进行干预。测定细胞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)及诱导型一氧化氮合酶(i NOS)的含量;测定一氧化氮(NO)的分泌水平;测定细胞内环磷酸腺苷(cAMP)水平及四唑盐(MTT)比色试验测定细胞活力。结果西洛他唑组及依达拉奉组与糖氧剥离模型组比较,LDH、MDA漏出量显著减少(P均<0.05),GSH-Px释放量明显升高(P均<0.05),nNOS、i NOS的水平及NO的分泌量显著下降(P均<0.05),细胞内cAMP水平明显升高(P均<0.05);细胞存活率显著提高(P均<0.05);西洛他唑与依达拉奉组比较,LDH、MDA漏出量及GSH-Px的释放量无差别,nNOS、i NOS和NO的水平明显降低(P均<0.05),细胞内cAMP水平显著升高(P<0.05);细胞存活率明显提高(P<0.05)。结论西洛他唑对培养大鼠皮层细胞在糖氧剥离损伤中具有保护作用,其作用机制可能通过抗氧化、降低nNOS及i NOS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 investigate the effects and possible mechanism of cilostazol on cultured cerebral cortical cells deprived of oxygen and glucose in rats in vitro.Methods The cortical cells of the neonatal Wistar rats were cultured in vitro,and then cell model of ischemia was established by oxygen-glucose deprivation（OGD） mimicking cerebral ischemia.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 6h OGD cell culture.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 OGD group,the concentrations of LDH,MDA,nNOS,iNOS and NO were significantly lower,the levels of GSH-Px and intracellular cAMP were obviously higher,and cells viability was marked increased in cilostazol group and edaravone group（all P0.05）.Compared with the edaravone group,no obvious difference was found in the concentrations of LDH,MDA and GSH-Px（all P0.05）,but the concentrations of nNOS,iNOS and NO were significantly lower（all P0.05） in the cilostazol group.And the intracellular cAMP levels and cells viability in the cilostazol group were significantly higher than those in the edaravone group（all P0.05）.Conclusions Our results show that cilostazol has the protective effects on cultured rat cortical cells in OGD.And the mechanism may be related with resisting oxidation,reducing levels of nNOS and iNOS which result in a reduction of NO secretion,and elevating intracellular cAMP levels.

关 键 词：皮层细胞 糖氧剥离 西洛他唑 保护作用 

分 类 号：R285.5[医药卫生—中药学]