细胞周期依赖激酶抑制剂Olomoucine对活化星形胶质细胞神经蛋白聚糖、短蛋白聚糖表达的抑制作用  

The expression of brevican and neurocan in the activity astrocytes and Inhibition effect of Olomoucine

作　　者：易善清[1] 侯超[2] 刘珍[3] 武衡[1] 

机构地区：[1]南华大学附属第一医院神经内科,湖南衡阳421001 [2]湖南省第二人民医院,湖南长沙410001 [3]南华大学机能实验室,湖南衡阳421001

出　　处：《中风与神经疾病杂志》2017年第6期504-507,共4页Journal of Apoplexy and Nervous Diseases

基　　金：衡阳市科技局(2015KJ43)

摘　　要：目的研究细胞周期依赖激酶抑制剂Olomoucine对星形胶质细胞活化后神经蛋白聚糖(Neurocan)、短蛋白聚糖(Brevican)表达的抑制作用。方法应用睫状神经营养因子(CNTF)刺激法建立体外培养星形胶质细胞的活化模型,实验分3组:对照组、活化组与抑制组。对照组:正常培养的星形胶质细胞;活化组:星形胶质细胞加入20 ng/ml睫状神经营养因子(CNTF)处理24 h;抑制组:星形胶质细胞用20 ng/ml睫状神经营养因子预处理24 h,加入100μmol/L Olomoucine。应用ELISA法检测不同时间点(12 h、24 h、48 h)各组细胞上清液中Neurocan、Brevican含量变化,半定量RT-PCR检测各组细胞GFAP mRNA及Neurocanm RNA、Brevican mRNA的表达变化。结果(1)活化组上清液中Neurocan、Brevican的含量在12 h、24 h、48 h随着时间的延长逐渐增加,与对照组比较差异有显著性(P<0.01);抑制组上清液中Neurocan、Brevican含量随着时间的延长较活化组明显降低,与对照组、活化组比较差异有显著性(P<0.01)。(2)活化组细胞GFAP mRNA、Neurocan mRNA、Brevican mRNA表达明显增加,与对照组比较差异有显著性(P<0.01);抑制组细胞GFAP mRNA、Neurocan mRNA、Brevican mRNA表达均明显下调,与活化组比较差异有显著性(P<0.01)。结论细胞周期依赖激酶抑制剂Olomoucine可抑制活化星形胶质细胞神经蛋白聚糖、短蛋白聚糖表达。Objective To study the inhibitory effect of cell cycle-dependent kinase inhibitor Olomoucine on the expression of neuronal glycoprotein（ Neurocan） and short protein oligosaccharides（ Brevican） after activation of astrocytes.Methods To establish the activation model of astrocytes cultured in vitro by using ciliary neurotrophic factor（ CNTF）stimulation method. The experiment were divided into three groups： the control group,the activation group and the suppression group. Astrocytes in the control group were cultured generally; the activation group were co-cultuted with 20 ng/ml CNTF,and in the suppression group were incubated 100 μmol/L olomoucine. Three groups of cells by different conditions deal with 12,24 and 48 hours later. The content of Neurocan and Brevican were examined by sites sandwich enzyme linked immunosorbent assay（ ELISA）. RT-PCR was used to detect the expression of glial fibrillary acidic protein（ GFAP） mRNA,Neurocanm RNA and Brevicanm RNA. Results（ 1） The content of Neurocan and Brevican in the supernatant of activation group was gradually increased with the duration of 12,24 and 48 h,and the difference was significant compared with the control group（ P〈 0. 01）. The content of Neurocan and Brevican in inhibition group was significantly decreased with the duration of the activation group,and the difference was significant compared with control group and activation group（ P〈 0. 01）.（ 2） The expression of GFAP mRNA,Neurocan mRNA and Brevican mRNA in activation group was significantly increased,and the difference was significant compared with the control group（ P〈 0. 01）. The expression of GFAP mRNA,Neurocan mRNA and Brevican mRNA decreased markedly,and the difference was significantly compared with activation group（ P〈 0. 01）. Conclusions Cell cycle-dependent kinase inhibitor Olomoucine inhibits the expression of activated stellate glial cell nerve protein glycoprotein and short protein.

关 键 词：星形胶质细胞 OLOMOUCINE 神经蛋白聚糖 短蛋白聚糖 

分 类 号：R741[医药卫生—神经病学与精神病学]