智脑胶囊调控PI3K/AKT/GSK-3β信号通路诱导阿尔茨海默病模型细胞增殖的研究  

Investigation of Zhinao Capsule induces proliferation of Alzheimer’s disease model cells by PI3K/AKT/GSK-3β signal pathway

作　　者：姜辉[1] 凡畅 汪永忠[2] 杨文明[3] JIANG Hui;FAN Chang;WANG Yong-zhong;YANG Wen-ming(Clinical Research and Experimental Center,The First Affiliated Hospital of Anhui University of Chinese Medicine,Anhui Hefei 230031,China;Department of Pharmacy,The First Affiliated Hospital of Anhui University of Chinese Medicine,Anhui Hefei 230031,China;Encephalopathy Center,The First Affiliated Hospital of Anhui University of Chinese Medicine,Anhui Hefei 230031,China)

机构地区：[1]安徽中医药大学第一附属医院临床研究实验中心,安徽合肥230031 [2]安徽中医药大学第一附属医院药学部,安徽合肥230031 [3]安徽中医药大学第一附属医院脑病中心,安徽合肥230031

出　　处：《中国医院药学杂志》2022年第24期2612-2617,共6页Chinese Journal of Hospital Pharmacy

基　　金：安徽省科技重大专项项目(编号:201903a07020016)。

摘　　要：目的:探讨智脑胶囊(Zhinao Capsule, ZNC)对阿尔茨海默病(Alzheimer’s disease, AD)体外细胞模型增殖的影响及其作用机制。方法:采用全反式维甲酸(all trans retinoic acid, ATRA)+冈田酸(okadaic acid, OA)刺激人神经母细胞瘤细胞(human neuroblastoma cell, SK-N-SH)建立AD体外模型,CCK8法筛选ZNC含药血清最佳作用浓度和时间;流式细胞术、RT-qPCR和Western blot法检测最佳浓度ZNC含药血清作用最佳时间后,对SK-N-SH细胞周期、凋亡及PI3K、AKT、GSK-3βmRNA、总蛋白和磷酸化蛋白表达的影响。结果:ATRA+OA刺激后,SK-N-SH细胞活力降低,细胞凋亡增加,p-PI3K、p-AKT和p-GSK-3β蛋白表达显著降低,GSK-3βmRNA和总蛋白表达明显升高;给予ZNC干预后,不仅可明显促进SK-N-SH细胞增殖,抑制其凋亡,还可显著升高p-PI3K、p-AKT和p-GSK-3β蛋白表达,降低GSK-3βmRNA和总蛋白表达。结论:智脑胶囊可促进AD模型细胞增殖,其机制可能与调控PI3K/AKT/GSK-3β信号通路有关。OBJECTIVE To explore the effect of Zhinao Capsule(ZNC)on cell proliferation of Alzheimer’s disease(AD)cell model in vitro and its possible molecular mechanism.METHODS Human neuroblastoma cells(SK-N-SH)were induced by all trans retinoic acid(ATRA)and stimulated by okadaic acid(OA)to establish AD cell model.The optimal concentration and time of ZNC drug containing serum were screened by CCK8 assay.Flow cytometry, RT-qPCR and Western blot assays were used to detect the effects of ZNC on the cell cycle, apoptosis and the mRNA,protein and phosphorylated protein expression of PI3 K,AKT and GSK-3β in SK-N-SH cells.RESULTS After ATRA induction and OA stimulation, SK-N-SH cell viability decreased, apoptosis increased, p-PI3 K,p-AKT and p-GSK-3β protein expression decreased significantly, GSK-3β mRNA and protein expression increased significantly.ZNC drug-containing serum not only significantly promoted SK-N-SH cell proliferation and inhibited apoptosis, but also significantly increased p-PI3 K,p-AKT,p-GSK-3β protein expression, decreased GSK-3β mRNA and protein expression.CONCLUSION ZNC can promote the proliferation of AD cells, and its mechanism may be related to the regulation of PI3 K/AKT/GSK-3β signal pathway.

关 键 词：智脑胶囊 阿尔茨海默病 人神经母细胞瘤细胞 磷脂酰肌醇-3-激酶/蛋白激酶B/糖原合成酶激酶3β信号通路 

分 类 号：R964[医药卫生—药理学]