基于Aβ25-35诱导的PC12细胞损伤模型研究定志小丸治疗阿尔兹海默病的作用机制及配伍机制  

Study on Compatibility Mechanism of Ding-Zhi-Xiao-Wan Formula in Treating Alzheimer's Disease Based on Aβ_(25-35) Induced PC12 Cells

作　　者：郑妍 刘舒[1] 宋凤瑞[1] 刘志强[1,3] ZHENG Yan;LIU Shu;SONG Feng-Rui;LIU Zhi-Qiang(National Center of Mass Spectrometry in Changchun&Jilin Province Key Laboratory of Chinese Medicine Chemistry and Mass Spectrometry&Chemical Biology Laboratory,Changchun Institute of Applied Chemistry, Chinese Academy of Sciences,Changchun 130022,China;University of Chinese Academy of Sciences,Beijing 100049,China;State Key Laboratory of Electroanalytical Chemistry,Changchun Institute of Applied Chemistry, Chinese Academy of Sciences,Changchun 130022,China)

机构地区：[1]中国科学院长春应用化学研究所,国家质谱中心(长春),吉林省中药化学与质谱重点实验室及化学生物学实验室,长春130022 [2]中国科学院大学,北京100049 [3]中国科学院长春应用化学研究所电分析化学国家重点实验室,长春130022

出　　处：《分析化学》2018年第11期1724-1731,共8页Chinese Journal of Analytical Chemistry

基　　金：国家自然科学基金项目(No.81530094)资助。

摘　　要：利用β-淀粉样蛋白片段Aβ_(25-35)诱导的PC12细胞损伤模型,从细胞凋亡及氧化应激两个通路研究定志小丸对Aβ_(25-35)引起的PC12神经细胞损伤的保护作用,并将定志小丸拆方为单味药及三味药,用以阐明定志小丸治疗阿尔兹海默病的作用机制、配伍机制以及主要活性药味。通过MTT和乳酸脱氢酶含量测定评估定志小丸及各味药对PC12细胞活力的影响,其中人参、茯苓效果最好,与模型组相比,可使细胞活力提高约30%。通过细胞凋亡、线粒体膜电位(MMP)、丙二醛(MDA)及还原型谷胱甘肽(GSH)含量测定,进一步从细胞凋亡及氧化应激两个通路研究定志小丸及各味药对Aβ_(25-35)诱导PC12细胞保护作用,研究结果表明,人参、茯苓效果最优,不仅可以显著抑制细胞凋亡,还可降低丙二醛(MDA)含量,减少膜脂过氧化;远志、石菖蒲可以通过提高细胞内还原型谷胱甘肽(GSH)含量进而抑制氧化应激损伤,且与人参、茯苓配伍后,可促进主要成分发挥药效,显著提高人参、茯苓的药效。人参、茯苓是定志小丸中的主要活性药味,远志、石菖蒲起辅助增强药效的作用。The protective effect of Ding-Zhi-Xiao-Wan(DZXW)on Aβ25-35 induced PC12 cells was investigated from two aspects of cell apoptosis and oxidative stress.And the formula was decomposed into single herbs and three herbs to clarify the mechanisms of action,compatibility mechanisms and main active ingredients of DZXW.The viability of PC12 cells was evaluated by MTT assay and lactate dehydrogenease content,ginseng and poria cocos played the key roles in enhancing cell viability.Compared with the model group,it could improve the cell viability about 30%.The protection mechanism of DZXW against Aβ25-35 induced PC12 cells injury on oxidative stress and cell apoptosis pathways were studied by flow cytometry,mitochondrial membrane potential(MMP),content of malondialdehyde(MDA)and glutathione(GSH)for the further research.Ginseng and poria could significantly inhibit the cell apoptosis,and also decreased MDA content to reduce the membrane lipid peroxidation.The effect of polygala radix(PR)and acorus tatarinowii rhizoma(ATR)could improve the content of GSH in PC12 cells to inhibit oxidative stress injuries,and the activities of ginseng and poria were improved after the compatibility with PR and ATR.Overall speaking,the compatibility of DZXW formula was scientific rationality.

关 键 词：定志小丸 拆方 配伍作用 AΒ25-35 PC12细胞 细胞凋亡 氧化应激 

分 类 号：R285.5[医药卫生—中药学]