Bcl-2和Bad参与GDNF协同雌二醇对多巴胺能神经细胞的保护作用  

Bcl-2 and Bad participate in the protective effect of GDNF acting in synerg acting in synergy with estradiol on dopaminergic neurons

作　　者：刘亚萍[1] 董富兴[2] 王晓舟[3] 高殿帅[2] 

机构地区：[1]徐州医学院形态学实验教学中心,徐州221002 [2]徐州医学院神经生物学实验中心,徐州221002 [3]徐州医学院机能学实验中心,徐州221002

出　　处：《神经解剖学杂志》2015年第1期44-48,共5页Chinese Journal of Neuroanatomy

基　　金：国家自然科学基金(30870797);徐州医学院院课题(2012KJ09)

摘　　要：目的：研究GDNF和雌二醇（E2）联合使用对6-羟基多巴胺（6-OHDA）所致的多巴胺能神经细胞损伤的影响并探讨其作用的机制.方法：新生2～3d的雄性SD大鼠行常规中脑脑片培养,6-OHDA诱导损伤,脑片分为8组：正常对照组、溶剂对照组、E2组、Wortmannin（PI3-K特异性阻断剂）＋E2组、GDNF组、Wortmannin＋GDNF组、GDNF＋E2组、Wortmannin＋GDNF＋E2组.免疫荧光染色观察黑质致密部（SNc）酪氨酸羟化酶（TH）的表达情况;Western Blot检测各组脑片中Bcl-2及Bad的表达.结果：免疫荧光染色结果显示,在GDNF＋E2组,TH的表达比单用GDNF或E2时显著升高,差异有统计学意义（P＜0.05）;阻断PD-K/Akt信号通路后,TH的表达与未阻断组相比降低,差异有统计学意义（P＜0.05）.Western Blot检测结果显示,在GDNF＋ E2组,Bcl-2的表达比单用GDNF或E2时升高,而Bad的表达比单用GDNF或E2时降低,差异均有统计学意义（P＜0.05）;阻断PI3-K/Akt信号通路后,Bcl-2的表达与未阻断组相比降低,而Bad的表达是升高的,差异有统计学意义（P＜0.05）.结论：GDNF和E2联合使用对6-OHDA所致的黑质多巴胺能神经细胞损伤有保护作用,其机制可能是通过调节凋亡相关蛋白Bcl-2和Bad的表达变化实现的.Objective： To study the effects and the mechanisms of GDNF and estraiol （E2） on dopaminergic（DA） neuronal which was injuried by 6-hydroxydopamine （6-OHDA）. Methods： The 2-3-day-old male SD rats underwent conventional midbrain slice cultures, and then the slices were divided randomly into eight groups after injuried by 6-OHDA ： norreal control group, experimental control group, E2 group, Wortmannin （PI3-K inhibitors） ＋ E2 group, GDNF group, Wortmannin ＋ GDNF group, GDNF ＋ E2 group, Wortmannin ＋ GDNF ＋ E2 group. The expressions of tyrosine hydroxylase （TH） in substantia nigra pars compacta （SNc） were observed by immunofluorescence. The Bcl-2 and Bad expressions in DA neurons were detected by Western Blot. Results： The immunofluorescence results showed, the expression of TH in GDNF ＋ E2 group was significantly higher than that in the GDNF group and E2 group （ P〈0. 05）. After blocking the P13-K/Akt pathway, the expressions of TH were decreased in the Wortmannin groups compared with those without blocking （P 〈0. 05 ）. The expression of Bcl-2 in GDNF ＋ E2 group was higher than that in the E2 group and GDNF group, while the difference was statistically significant （ P 〈 0.05 ）. The expression of Bad in GDNF ＋ E2 group was lower than that in GDNF group and E2 group （P 〈 0.05）. After blocking PI3-K/Akt pathway, the expressions of Bcl-2 were decreased in the Wortmannin groups while the expressions of Bad were increased, which were compared with groups without blocking （P 〈 0.05）. Conclusion ： GDNF in combination with E2 have protective effects on dopaminergic neurons injured by 6-OHDA, which may be mediated by regulating Bcl-2 and Bad.

关 键 词：雌激素 多巴胺 GDNF BCL-2 BAD 大鼠 

分 类 号：R741[医药卫生—神经病学与精神病学]