硝酸甘油对体外培养三叉神经节星形胶质细胞的损伤作用  

Injured effect of GTN on primary cultured trigeminal ganglion astroglia in vitro

作　　者：崔智威[1] 曾照芳[1] 陈力学[2] 周冀英[2] 申崇标[1] 

机构地区：[1]重庆医科大学医学检验系临床检验诊断学教育部重点实验室重庆市重点实验室,重庆400016 [2]重庆医科大学附属第一医院重庆市神经病学重点实验室,重庆400016

出　　处：《激光杂志》2010年第5期63-65,共3页Laser Journal

基　　金：重庆市科委自然科学基金资助项目(CSTC;2007BB5287);重庆市卫生局医学科研资助项目(2009-2-357)

摘　　要：目的:硝酸甘油(glyceryl trinitrate或nitroglycerin,GTN)对于体外培养的三叉神经节星形胶质细胞损伤作用,为偏头痛发病机制研究提供依据。方法:体外纯化培养大鼠三叉神经节星形胶质细胞,采用胶质纤维酸性蛋白(GFAP)免疫荧光法鉴定。不同浓度的硝酸甘油作用于星形胶质细胞后,应用Alarm blue检测细胞活力,并筛选出可以影响细胞活力的适宜刺激的浓度;适宜刺激后用Fluo-3AM为细胞内钙离子荧光指示剂,激光共聚焦显微镜测定各组星型胶质细胞内钙离子浓度的变化;RT-PCR检测IL-1β、TNF-αmRNA表达。结果:GFAP免疫荧光法显示培养的星型胶质细胞阳性率达95%左右;0.55mmol/L、1.1mmol/L和2.2mmol/L三种浓度硝酸甘油刺激星型胶质细胞后活力明显降低(P<0.05),但从细胞活力和形态上观察0.55mmol/L硝酸甘油刺激最佳;0.55mmol/L硝酸甘油刺激后细胞内钙略降低,IL-1β、TNF-αmRNA表达增高(P<0.05)。结论:硝酸甘油对于体外培养的星型胶质细胞有明显的损伤作用,其机制可能与炎症因子相关。Objective：To study the injured effects induced by GTN on primary cultured trigeminal ganglion astroglia.Methods：Rat astroglia were cultured and identified by glial fibrillary acid protein（GFAP）;Alarm blue assay detected cell viability after different concentrations of GTN acts on astrocytes,and filtered that can affect the viability of the appropriate concentration of stimulating;The change ofin astroglia after the stimulation of GTN was detected by means of a confocal laser scanning microscope using the（Ca^2＋） indicator Fluo-3AM;The expression of IL-Iβ,TNF-α mRNA were detected by RT-PCR.Results：Cells can expressed GFAP,and astroglia activity decreased with 0.55,1.1 and 2.2mmol/L GTN stimulating,but according to the viability and modality of the cells,0.55mmol/L GTN was quite suited;The change of（Ca^2＋）in reduced and the expression of IL-1β,TNF-α mRNA in astroglia increased. Conclusion：GTN can demaged primary cultured trigeminal ganglion astroglia that were related with inflammatory factor relased.

关 键 词：硝酸甘油 偏头痛 星形胶质细胞 IL-1Β TNF-α 

分 类 号：R477[医药卫生—护理学]