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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 investigate the protective effect of dopamine (DA) on apoptosis of pituitary adenoma GH4 cells and glutathione (GSH) on DA induced apoptosis. Methods In this experiment, the apoptotic effect of DA and the protective effect of GSH were studied in three parts: ①The experimental group was divided into blank control group and DA treatment group, the effect of different concentration and time of DA on the growth of GH4 cells was observed in vitro; ②The experimental group was divided into blank control group, DA group and DA combined with DA D2 receptor antagonist group to observe the role of D2 receptor in apoptosis; ③The experiment was divided into blank control group, DA group, GSH treatment group, PI staining were observed in three groups of cell apoptosis Western blot was used to detect the expression of Bcl-2 and PARP-1. Results DA induced apoptosis in GH4 cells in a time-and dose-dependent manner, selective D2 receptor antagonists did not block apoptosis. After GSH-treated GH4 cells, PI staining showed that the number of apoptotic cells was significantly lower than that in DA group, Western blot Bcl-2 expression increased, PARP-1 expression decreased. CONCLUSION DA induces apoptosis of GH4 cells through intracellular action. Selective D2 receptor antagonist can not block apoptosis. GSH may protect DA-induced apoptosis of GH4 cells, which may be related to up-regulation of Bcl-2, up-regulation of PARP- 1 drop related.