丁基苯酞对氧糖剥夺/复氧糖诱导的星形胶质细胞中caspase-3表达的影响  

Effect of butylphthalide on the expression of caspase-3 in astrocytes induced by oxygen-glucose deprivation and reoxygenation

作　　者：殷闯[1] 牛草原[1] 胡亚男[2] 

机构地区：[1]新乡医学院第三附属医院神经内科,河南新乡453003 [2]新乡医学院三全学院组织学与胚胎学教研室,河南新乡453003

出　　处：《新乡医学院学报》2017年第5期365-368,373,共5页Journal of Xinxiang Medical University

摘　　要：目的探讨丁基苯酞对氧糖剥夺/复氧糖诱导的星形胶质细胞中caspase-3表达的影响。方法将原代培养、鉴定的Sprague-Dawley大鼠星形胶质细胞随机分为对照组、模型组和实验组,各组细胞经高糖达尔伯克改良伊格尔培养基(DMEM)培养48 h,细胞贴壁并伸出突触后实验组及模型组更换DMEM无糖培养基进行氧糖剥夺,对照组仍使用DMEM高糖培养基培养,3组细胞放入培养箱中孵育6 h后,实验组更换为含100μmol·L^(-1)丁基苯酞的DMEM高糖培养基,模型组更换为DMEM高糖培养基进行氧糖剥夺/复氧糖,对照组更换新的DMEM高糖培养基,继续培养24 h。四甲基偶氮唑盐(MTT)法检测各组细胞活力,免疫荧光及免疫印迹法检测星形胶质细胞中caspase-3表达,酶联免疫吸附试验检测细胞培养液中乳酸脱氢酶(LDH)表达。结果 MTT法结果显示,对照组星形胶质细胞活力高于实验组和模型组(P<0.05),实验组星形胶质细胞活力显著高于模型组(P<0.05)。免疫荧光染色结果显示,对照组星形胶质细胞内荧光信号最弱;模型组星形胶质细胞内荧光信号最强;实验组星形胶质细胞内荧光信号强度较模型组降低,但高于对照组(P<0.05)。免疫印迹检测结果显示,模型组星形胶质细胞在氧糖剥夺/复氧糖后caspase-3表达相对最高,实验组次之,对照组最低,各组间两两比较差异均有统计学意义(P<0.05)。氧糖剥夺/复氧糖24 h后,实验组细胞培养液中LDH水平高于对照组(P<0.05),模型组细胞培养液中LDH水平显著高于实验组(P<0.05)。结论丁基苯酞可以改善氧糖剥夺/复氧糖引起的星形胶质细胞损伤。Objective To study the effect of butylphthalide on the expression of caspase-3 in astrocytes induced by oxygen-glucose deprivation and reoxygenation （OGD/R）.Methods Astrocytes of Sprague-Dawley rats were cultured and identified and randomly divided into control group,model group and experimental group,the cells in each group were cultured by high glucose Dulbecco＇s modified Eagle＆#39;s medium（DMEM） for 48 h.The medium in the model group and experimental group was replaced by sugar-free DMEM after cell adhesion and protruding synapse,while the cells in control group were cultured in the quondam medium.After the cells in the three groups were incubated for 6 h in incubator,the medium in experimental group was replaced by high glucose DMEM with 100 pmol ＆#183; L-1 butylphthalide,and the medium of model group was replaced by high glucose DMEM,while the medium of the control group was replaced by new high glucose DMEM.All the cells in the three groups were cultured for 24 h sequentially.The cell activity was detected by methyl thiazolyl tetrazolium （MTT） method.The expression of caspase-3 was measured by immunofluorescence and immunoblotting.The level of lactic dehydrogenase （LDH） was detected by enzyme linked immunosorbent assay.Results MTT results showed that the astrocyte activity of the control group was higher than that of the experimental group and model group （P 〈 0.05）,while the astrocyte activity of the experimental group was higher than that of the model group （P 〈 0.05）.Immunofluorescence staining showed that the fluorescence signal in the astrocyte of the model group was the strongest.The fluorescence intensity in the experimental group was lower than that in the model group,but higher than that in the control group （P 〈 0.05）.Western blot analysis showed that the expression of caspase-3 in astrocytes after OGD/R in the model group was relatively the highest,it in the experimental group took second place,while it in the control group was the lowest,there was signific

关 键 词：CASPASE-3 丁基苯酞 星形胶质细胞 氧糖剥夺/复氧糖 

分 类 号：R965[医药卫生—药理学]