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目的探讨黄芪提取物(EA)对Aβ_(25-35)所致海马神经元损伤的保护作用及其作用机制。方法分离孕18 d的胎鼠的海马神经细胞进行体外培养,实验分为3组:对照组、模型组和黄芪组。对照组:只加入无B27神经生长因子的神经基础培养基(NB)原液培养24 h;模型组:加入无B27神经生长因子的NB原液,再加入终浓度为10μmol/L Aβ_(25-35)培养24 h;黄芪组:加入无B27神经生长因子的NB原液,先加入终浓度为黄芪提取物20 mg/L培养4 h,再加入终浓度为10μmol/L Aβ_(25-35)培养20 h。在倒置显微镜下观察各组海马神经细胞形态学变化;采用乳酸脱氢酶(LDH)法和3-(4,5-甲基噻唑-2)-2,5-苯基的氮唑溴盐(MTT)法检测细胞活性;Western blot检测各组海马神经细胞Bcl-2和Bax蛋白的表达情况。结果模型组的海马神经细胞活性明显下降,细胞出现凋亡的典型的形态学特征,Bcl-2蛋白的表达水平下降而Bax蛋白的表达水平升高,与对照组比较,差异有统计学意义(P<0.05)。黄芪组海马神经细胞活性提高、凋亡的细胞数减少,Bcl-2蛋白的表达水平升高而Bax蛋白的表达水平下降,与模型组比较,差异有统计学意义(P<0.05)。结论黄芪提取物对Aβ_(25-35)所诱导的海马神经元损伤具有一定的保护作用,可能与其降低海马神经细胞Bcl-2蛋白的表达水平升高Bax蛋白的表达水平有关。
Objective To investigate the protective effect of astragalus extract (EA) on hippocampal neuron injury induced by Aβ_ (25-35) and its mechanism. Methods The fetal rat hippocampal neurons isolated from pregnant mice were cultured in vitro. The experiment was divided into three groups: control group, model group and Astragalus group. The control group: NB medium containing only B27 NGF was cultured for 24 h; model group: NB medium without B27 NGF was added, and then added to a final concentration of 10 μmol / L Aβ_ (25-35) The cells were cultured for 24 h. The Astragalus membranaceus group was treated with NB stock solution without B27 nerve growth factor. The final concentration of astragalus extract 20 mg / L was added for 4 h, then the cells were incubated with 10 μmol / L Aβ 25-35 for 20 h . Morphological changes of hippocampal neurons were observed under an inverted microscope. Lactate dehydrogenase (LDH) method and azolesbromide salt of 3- (4,5-methylthiazol-2) MTT assay was used to detect the cell viability. Western blot was used to detect the protein expression of Bcl-2 and Bax in each group. Results The neuronal activity of hippocampus in model group was significantly decreased. The typical morphological features of apoptosis were observed in the model group. The expression of Bcl-2 protein was decreased and the expression of Bax protein was increased. Compared with the control group, the difference was statistically significant ( P <0.05). Compared with model group, the activity of hippocampal neurons in astragalus increased, the number of apoptotic cells decreased, the expression of Bcl-2 protein increased and Bax protein decreased (P <0.05). Conclusion Astragalus extract can protect hippocampal neurons induced by Aβ 25-35, which may be related to the decrease of Bcl-2 protein expression and the expression of Bax protein in hippocampal neurons.