基于Nrf2/Keap1信号通路人参皂苷CK靶向调节Aβ分子机制研究  

Research on ginsenosides CK targeted regulating Aβ molecular to protect the damaged neurons based on Nrf2/Keap1 signaling pathways

作　　者：杨擎[1] 隋欣[1] 石晓征[1] 李晓华[1] 韩冬[1] 林嘉楠[1] 刘英娜[1] 张慧媛[1] 阚默 修志儒 李娜[1] 曲晓波[1] YANG Qing;SUI Xin;SHI Xiao-zheng;LI Xiao-hua;HAN Dong;LIN Jia-nan;LIU Ying-na;ZHANG Hui-yuan;KAN Mo;XIU Zhi-ru;LI Na;QU Xiao-bo(Changchun University of Chinese Medicine, Changchun 130117, China)

机构地区：[1]长春中医药大学,长春130117

出　　处：《中华中医药杂志》2018年第5期1840-1845,共6页China Journal of Traditional Chinese Medicine and Pharmacy

基　　金：国家自然科学基金项目(No.81704001);吉林省教育厅"十三五"科学技术项目(No.JJKH20170713KJ);吉林省科技厅自然科学基金项目(No.20160101229JC)~~

摘　　要：目的:基于β-淀粉样蛋白(Aβ)靶点探讨人参皂苷Compound K(CK)对受损神经元保护作用及下游Nrf2/Keap1信号通路调控作用。方法:通过Open SPR技术检测CK在体外对Aβ配体的结合常数、解离常数及解离平衡常数;SDS-PAGE实验检测CK对Aβ寡聚体解聚的影响。在细胞水平检测CK对Aβ损伤SH-SY5Y细胞存活率的影响,利用DCFH-DA探针检测受损神经元活性氧簇(ROS)表达水平。蛋白水平通过Western blot检测与调节Aβ相关激酶BACE1、PS1、IDE的表达情况,并检测阿尔茨海默病病理特征性蛋白Aβ1-42表达及其下游Nrf2/Keap1信号通路蛋白表达的水平。结果:CK能增加Aβ的解聚能力,提高Aβ损伤SH-SY5Y细胞存活率(P<0.05),降低ROS产生(P<0.05),抑制Aβ、BACE1、PS1的表达(P<0.05),促进IDE的表达(P<0.05),激活Nrf2/Keap1信号通路的表达(P<0.05)。结论:CK能靶向调节Aβ并促进Aβ寡聚体解聚,抑制ROS的产生,减轻氧化应激反应,其机制可能与靶向亲和Aβ寡聚体使其解聚并减少ROS的表达,进而激活Nrf2/Keap1通路有关。Objective： To study the protective effect of Compound K（CK） on damaged neurons and the molecular mechanism of Nrf2/Keap1 signaling pathway based on Aβ target. Methods： The binding constants, dissociation constants and dissociation equilibrium constants of CK in vitro were determined by Open SPR. The effect of CK on depolymerization of Aβoligomers was researched by SDS-PAGE. The effects of CK on the survival rate of SH-SY5 Y cells were detected, and the ROS of injured neurons was detected by DCFH-DA at the cellular level. Western blotting was used to detect the expression of BACE1, PS1, IDE, the pathological characteristic protein of Alzheimer Aβ1-42, and the protein of Nrf2/Keap1 signal pathway. Results： CK could increase depolymerzation ability of Aβ, improve the survival rate of SH-SY5 Y cells damaged by Aβ（P〈0.05）, reduce the generation of ROS（P〈0.05）, inhibit the expression of Aβ, BACE1 and PS1（P〈0.05）, promote the expression of IDE（P〈0.05）, and activate Nrf2/Keap1 signal pathway（P〈0.05）. Conclusion： CK could target adjust and promote depolymerztion of Aβoligomers, inhibite the production of ROS, and reduce oxidative stress. The mechanism might be related with the depolymerization and down-regulation of oligomers neuron Aβ expression which decreased the expression of ROS, and then further activate Nrf2/Keap1 pathway.

关 键 词：人参皂苷compound K Β-淀粉样蛋白 Nrf2/Keap1 氧化应激 阿尔茨海默病 

分 类 号：R285[医药卫生—中药学]