[Effects of Huangpu Tongqiao capsule on apoptosis of Alzheimer's disease cell model].
[Effects of Huangpu Tongqiao capsule on apoptosis of Alzheimer's disease cell model].
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DOI：
10.19540/j.cnki.cjcmm.20180403.001
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发表时间：
2018-06-01
期刊：
影响因子：
--
通讯作者：
Xie, Dao-Jun
中科院分区：
文献类型：
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作者：
Cai, Biao;Ye, Shu;Xie, Dao-Jun
海马神经元丢失是阿尔茨海默病(AD)的主要病理特征之一，与海马神经元的凋亡有关。黄埔通窍胶囊用于治疗阿尔茨海默病，但其作用机制尚不清楚。本研究通过观察黄埔通窍胶囊含药血清对Aβ₂₅₋₃₅诱导的原代培养海马神经元细胞凋亡的影响，探讨黄埔通窍胶囊治疗AD的神经保护作用机制。原代培养海马神经元，MAP-2免疫荧光染色鉴定，倒置显微镜观察细胞生长状态。采用血清药理学方法制备黄埔通窍胶囊含药血清。采用四甲基偶氮唑盐比色法测定黄埔通窍胶囊含药血清的最佳血药浓度范围，以及建立AD模型的最佳Abeta浓度。用Aβ₂₅₋₃₅诱导原代培养的海马神经元AD细胞模型后，用四甲基偶氮唑盐比色法检测细胞存活率，流式细胞仪检测细胞凋亡率，Western印迹法检测bax、细胞色素C和半胱氨酸天冬氨酸蛋白酶3的蛋白表达。结果显示：原代培养的海马神经元培养成功，第7天细胞成熟，与正常组相比，AD细胞模型组海马神经元存活率降低，海马神经元凋亡率增加，Bax、CytC和caspase-3蛋白表达增加(P
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 Abeta₂₅₋₃₅ 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 Abeta concentration for establishing the AD model. After primary cultured hippocampal neurons AD cell model was induced by Abeta₂₅₋₃₅, 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