黄蒲通窍胶囊对阿尔茨海默病细胞模型细胞凋亡的影响  

Effects of Huangpu Tongqiao capsule on apoptosis of Alzheimer’s disease cell model

作　　者：蔡标[1,2] 叶树 王艳[1,2] 汪婷婷[1,2] 王靓 江爱娟[1,2] 方正清 申国明[1,2] 谢道俊 CAI Biao;YE Shu;WANG Yan;WANG Ting-ting;WANG Liang;JIANG Ai-juan;FANG Zheng-qing;SHEN Guo-ming;XIE Dao-jun(College of httegrated Chinese and Western Medicine,Anhui University of Chinese Medicine.Hefei 230012,China;Institute of Integrated Chinese and Western Medicine,Anhui Academy of Chinese Medicine,Hefei 230012,China;First Affiliated Hospital,Aahui University of Chinese Medicine.Hefei 230031,China)

机构地区：[1]安徽中医药大学中西医结合学院,安徽合肥230012 [2]安徽省中医药科学院中西医结合研究所,安徽合肥230012 [3]安徽中医药大学第一附属医院,安徽合肥230031

出　　处：《中国中药杂志》2018年第11期2378-2383,共6页China Journal of Chinese Materia Medica

基　　金：国家自然科学基金项目(81574040);安徽省高校优秀青年人才支持计划重点项目(gxyq ZD2018053)

摘　　要：海马神经细胞丢失是阿尔茨海默病（AD）的主要病理特征之一,其发生与海马神经细胞凋亡有关。黄蒲通窍胶囊用于治疗AD,但其作用机制尚未明确。该实验通过观察黄蒲通窍胶囊含药血清对Aβ25-35诱导的原代培养海马神经元AD细胞模型神经损伤的保护作用及细胞凋亡的影响,探讨黄蒲通窍胶囊治疗AD的神经保护作用机制。原代培养海马神经元,倒置显微镜观察细胞生长状态,MAP-2免疫荧光法鉴定原代培养的海马神经元。运用血清药理学方法制备黄蒲通窍胶囊含药血清,MTT法筛选黄蒲通窍胶囊含药血清最佳浓度范围和Aβ25-35造模浓度。用Aβ25-35诱导建立AD细胞模型后,给予不同浓度的黄蒲通窍胶囊含药血清继续干预,采用MTT法检测细胞存活率,流式细胞术检测细胞凋亡率,Western blot法检测细胞内Bax,Cyt C,caspase-3蛋白表达。结果显示,原代培养海马神经元成功,且海马神经元在培养至第7天时生长成熟;与正常组比较,模型组海马神经元细胞存活率下降,细胞凋亡率增加,Bax,Cyt C,caspase-3蛋白表达增加（P〈0.05,P〈0.01）;与模型组比较,黄蒲通窍胶囊含药血清组细胞存活率明显升高,细胞凋亡率减少,Bax,Cyt C,caspase-3蛋白表达明显降低（P〈0.05,P〈0.01）。结果表明,黄蒲通窍胶囊含药血清对Aβ25-35诱导的原代培养海马神经元损伤具有保护作用,可抑制海马神经元细胞凋亡从而起到治疗AD的作用。The loss of hippocampal neurons is one of the main pathological features of Alzheimer＇s disease（ AD）,which is related to the apoptosis of hippocampal neurons. Huangpu Tongqiao capsule is used for the treatment of AD,but the underlying mechanism is still unclear. This study is to investigate the mechanism of neuroprotective effect of Huangpu Tongqiao capsule in the treatment of AD,through observing the effect of Huangpu Tongqiao capsule containing serum on cell injury of primary cultured hippocampal neurons induced by Aβ25-35 via inhibiting the cell apoptosis. Primary cultured hippocampal neurons were cultured and identified by MAP-2 immunofluorescence staining,and cell growth state was observed by inverted microscope. The Huangpu Tongqiao capsule containing serum was prepared using the method of serum pharmacology. MTT assays were used to measure the optimum concentration range of Huangpu Tongqiao capsule containing serum,and optimum Aβ concentration for establishing the AD model. After primary cultured hippocampal neurons AD cell model was induced by Aβ25-35,cell survival rate was detected by MTT,cell apoptosis rate was assayed by flow cytometry,and protein expressions of Bax,Cyt C and caspase-3 were determined by Western blot analysis. The results showed that the primary cultured hippocampal neurons were cultured successfully,and cells grew mature at seventh days; Compared with normal group,the survival rate of hippocampal neurons in AD cell model group was decreased,the apoptosis rate of hippocampal neurons was increased,and the protein expressions of Bax,Cyt C and caspase-3 were increased（ P 0. 05,P 0. 01）; Compared with AD cell model group,the survival rate of hippocampal neurons in Huangpu Tongqiao capsule containing serum group was increased,the apoptosis rate of hippocampal neurons was decreased,and the protein expressions of Bax,Cyt C and caspase-3 were decreased（ P 0. 05,P 0. 01）. These findings suggest that Huangpu Tongqiao capsule containing serum has a neuroprotective effect on cell

关 键 词：黄蒲通窍胶囊 阿尔茨海默病 原代培养海马神经元 神经保护 细胞凋亡 

分 类 号：R285.5[医药卫生—中药学]