谷胱甘肽对多巴胺诱导的GH4细胞凋亡的保护作用  

Glutathione protects GH4 pituitary lactotrope tumor cells from apoptosis induced by dopamine

作　　者：王晗[1] 李书鹏[2] 姜玉华[1] 刘芳[2] 

机构地区：[1]山东大学第二医院肿瘤防治中心,济南250033 [2]多伦多大学成瘾与精神卫生中心神经分子生物学实验室

出　　处：《山东大学学报（医学版）》2011年第11期48-52,73,共6页Journal of Shandong University:Health Sciences

摘　　要：目的探讨多巴胺(DA)诱导垂体瘤GH4细胞凋亡及谷胱甘肽(GSH)对DA诱导细胞凋亡的保护作用机制。方法本实验通过3部分探讨DA的凋亡作用及GSH的保护作用:①实验分空白对照组及DA用药组,体外观察不同浓度、时间DA对GH4细胞生长的影响;②实验分空白对照组、DA组、DA联合DA D2受体拮抗剂组,观察D2受体在细胞凋亡中的作用;③实验设空白对照组、DA组、GSH用药组,PI染色分别观察3组细胞的凋亡情况,Western blot检测Bcl-2及PARP-1的表达。结果 DA诱导的GH4细胞凋亡呈浓度-时间依赖性,选择性D2受体拮抗剂不能阻断细胞凋亡,经GSH处理GH4细胞后,PI染色显示凋亡细胞数明显低于DA组,Westernblot示Bcl-2表达增加,PARP-1表达下降。结论 DA通过细胞内作用诱导GH4细胞凋亡,选择性D2受体拮抗剂不能阻断细胞凋亡,GSH对DA诱导的GH4细胞凋亡有明显的保护作用,可能与Bcl-2上调、PARP-1下降有关。Objective To explore mechanisms of dopamine（DA） inducing GH4 cell apoptosis and glutathione（GSH）protecting GH4 cells from apoptosis induced by DA.Methods ① GH4 pituitary cells were treated with DA at 0,100,300 and 500μmol/L for 24h,then treated with DA at 500μmol/L for 0,1,3,5,12 and 24h to select the appropriate concentration and time.② Then GH4 cells were treated with raclopride（a D2 receptor antagonist,Rac）and GSH to explore the effects of Rac and GSH on apoptosis.③Apoptotic cells were counted by an inverted phase contrast microscope.Morphological appearance was observed by PI labeling,and expressions of Bcl-2 and PARP-1 were detected by Western blot.Results DA induced concentration-and time-dependent GH4 cell apoptosis.A selective D2 receptor antagonist could not block the cytotoxic effect.PI revealed that exposure to GSH（1mmol/L） for 1h prior to the DA treatment attenuated DA-induced apoptosis.Western blot showed up-regulation of Bcl-2 and down-regulation of PARP-1.Conclusion DA exerts cytotoxic effects on GH4 cells mainly through auto-oxidation in the intracellular space.A selective D2 receptor antagonist cannot block DA-induced apoptosis,while GSH can block it,which may be relevant to regulation of Bcl-2 and PARP-1.

关 键 词：多巴胺 GH4细胞 细胞凋亡 谷胱甘肽 基因 BCL-2 基因 PARP-1 

分 类 号：R739.41[医药卫生—肿瘤]