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【目的】探讨原儿茶酸(PCA)在阿尔茨海默病(AD)细胞模型中的保护作用及机制。【方法】采用免疫荧光法鉴定β淀粉样蛋白(Aβ_(1-42))纤维聚体,将Aβ_(1-42)作用于PC12细胞,并于不同时间点(0、3、6、9、12、24 h)观察细胞形态,采用四甲基偶氮唑盐(MTT)法检测细胞存活率以确定造模条件,并用MTT法检测原儿茶酸对PC12细胞的毒性条件。采用预保护的给药方式,检测不同浓度的原儿茶酸对PC12细胞的干预作用。采用Western-blot法检测自噬相关标记蛋白Beclin1的表达水平,以探究原儿茶酸的保护作用机制。【结果】显微镜下观察到12、24 h时,Aβ_(1-42)白色团块聚集基本稳定,无明显差异,MTT检测细胞损伤率均达40%,由此确定12 h和24 h均可做为AD的造模条件。原儿茶酸在200~800μmol/L浓度作用24 h的条件下,细胞活力显著上升(P<0.01)。Western-blot结果显示:原儿茶酸可显著增加Beclin1的表达水平。【结论】原儿茶酸有助于改善Aβ_(1-42)诱导的PC12细胞毒性,其机制可能与增加自噬水平有关。
【Objective】 To investigate the protective effect and mechanism of protocatechuic acid (PCA) in Alzheimer’s disease (AD) cell model. 【Method】 Aβ 1-42 was identified by immunofluorescence staining and Aβ 1-42 was applied to PC12 cells at different time points (0, 3, 6, 9, 12,24 h). Cell viability was determined by MTT assay to determine the conditions of the model. MTT assay was used to detect the toxicity of protocatechuic acid on PC12 cells. Preventive administration was used to detect the effects of different concentrations of protocatechuic acid on PC12 cells. The expression of autophagy-related Beclin1 protein was detected by Western-blot to explore the protective mechanism of protocatechuic acid. 【Result】 The results showed that the aggregation of Aβ_ (1-42) white pellets was basically stable at 12 and 24 h under microscope. There was no significant difference between the two groups. The cell damage rates of MTT assay reached 40% As the modeling conditions of AD. Protocatechuic acid increased cell viability at 200 ~ 800μmol / L for 24 h (P <0.01). Western-blot results showed that protocatechuic acid can significantly increase the expression of Beclin1. 【Conclusion】 Protocatechuic acid helps to improve Aβ 1-42-induced PC12 cytotoxicity, which may be related to the increase of autophagy.