目的 探究当归芍药散对β淀粉样蛋白（amyloidβ-protein，Aβ）诱导的胆碱能神经元损伤的保护作用及相关机制。方法 以30μmol/L Aβ_25-35诱导基底前脑胆碱能神经元损伤模型，给予2.5%、5.0%和10.0%当归芍药散含药血清进行干预，采用MTT法检测细胞存活率；TUNEL法和碘化丙啶（PI）/Hoechst荧光双染检测神经元凋亡情况；采用胆碱乙酰转移酶（choline acetyltransferase，ChAT）/磷酸化环磷腺苷效应元件结合蛋白（phosphorylated cAMP-response element binding protein，p-CREB）和ChAT/剪切型半胱氨酸天冬氨酸蛋白酶-3（cleaved cystein-asparate protease-3，cleaved Caspase-3）荧光双染检测当归芍药散对胆碱能神经元的抗凋亡情况；采用Western blotting法检测CREB/B淋巴细胞瘤2（B-cell lymphoma 2，Bcl-2）信号通路相关蛋白的表达情况。结果 5.0%和10.0%当归芍药散含药血清可显著增加Aβ诱导的胆碱能神经元存活率（P＜0.01），改善Aβ诱导的胆碱能神经元凋亡（P＜0.01），降低Caspase-3活化水平（P＜0.01）；10%当归芍药散含药血清能逆转Aβ诱导p-CREB水平的降低（P＜0.01），增加抗凋亡蛋白Bcl-2的表达（P＜0.05）。结论 当归芍药散可能通过CREB/Bcl-2信号通路改善Aβ诱导的胆碱能神经元凋亡。

Objective To study the protective effect and mechanism of Danggui Shaoyao San (当归芍药散, DSS) on amyloid β-protein (Aβ)-induced cholinergic neuron injury. Methods Basal forebrain neuron injury model was induced by 30 μmol/L Aβ_25-35, 2.5%, 5.0% and 10.0% DSS containing serum were given for intervention, MTT assay was used to detect cell survival rate; TUNEL and PI/Hoechst fluorescence staining were used to detect neuronal apoptosis; Choline acetyltransferase (ChAT)/phosphorylated cAMP-response element binding protein (p-CREB) and ChAT/cleaved cystein-asparate protease-3 (Caspase-3) fluorescence double staining were used to detect the anti-apoptotic effect of DSS on cholinergic neurons; Western blotting was used to detect the expressions of CREB/B-cell lymphoma 2 (Bcl-2) signaling pathway related proteins.Results DSS serum (5.0% and 10.0%) significantly increased the cell survival rate of Aβ-injured cholinergic neurons (P< 0.01), improved Aβ-induced apoptosis (P< 0.01) and decreased activation level of Caspase-3 (P< 0.01). DSS serum (10%) reversed Aβ-induced decrease in p-CREB level (P< 0.01) and increased expression of anti-apoptotic protein Bcl-2 (P< 0.05). Conclusion DSS may ameliorate Aβ-induced apoptosis of cholinergic neurons through CREB/Bcl-2 signaling pathway.

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国家自然科学基金资助项目（81774129）；国家自然科学基金资助项目（82004184）；湖南省教育厅科研基金重点项目（21A0241）；2019年湖湘高层次人才聚集工程-创新人才（2019RS1064）；2021年度湖南省大学生创新创业训练计划项目（S202110541049）