五味子乙素靶向调节Aβ及下游NF-κB/TNF-α信号通路保护受损神经元的分子机制  

Study on neuroprotective effect of schisandrin B by regulating the Aβ and its downstream NF-κB/TNF-α pathway

作　　者：杨擎[1] 李娜[1] 隋欣[1] 李晓华[1] 石晓征[1] 刘英娜[1] 林嘉楠[1] 张慧媛[1] 阚默 修志儒 曲晓波[1] 

机构地区：[1]长春中医药大学,长春130117

出　　处：《中华中医药杂志》2017年第5期2064-2069,共6页China Journal of Traditional Chinese Medicine and Pharmacy

基　　金：2016年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)靶点探讨五味子乙素(Sch B)对受损神经元的保护作用及下游核因子κB(NF-κB)/肿瘤坏死因子(TNF-α)信号通路调节的分子机制。方法:本研究通过Open SPR技术检测Sch B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Sch B对Aβ寡聚体解聚的影响。在细胞水平检测Sch B对Aβ损伤SH-SY5Y细胞增殖率的影响,FACS检测受损神经元凋亡情况。蛋白水平通过Western blot检测线粒体凋亡途径相关蛋白Bcl-XL、Caspase-3的表达情况,并检测阿尔茨海默症病理特征性蛋白Aβ1-42、p-Tau表达及其下游NF-κB/TNF-α信号通路蛋白表达的水平。结果:Sch B能促进Aβ的解聚,增加Aβ损伤SHSY5Y细胞的增殖率,抑制受损SH-SY5Y细胞凋亡,增加Bcl-XL的表达和降低Caspase-3的表达,抑制Aβ1-42和p-Tau的表达水平,下调NF-κB/TNF-α信号通路的表达水平。结论:Sch B能靶向与Aβ配体亲和并有效促进Aβ寡聚体解聚,抑制神经元的凋亡,其机制可能与靶向亲和Aβ配体使其寡聚体解聚并下调神经元Aβ的表达,抑制Tau蛋白高度磷酸化,进而下调NF-κB/TNF-α通路有关。Objective: To study on neuroprotective effect of schisandrin B by regulating the Aβ and its downstream NF-κB/TNF-α pathway. Methods: The binding constants, dissociation constants and dissociation equilibrium constants of Sch B in vitro were determined by Open SPR. The effect of Sch B on depolymerization of Aβ oligomers was determined by SDS-PAGE. The effects of Sch B on the proliferation of SH-SY5 Y cells were detected, and the apoptosis of damaged neurons was detected by FACS at the cellular level. Western blot was used to detect the protein expressions involving the mitochondrial apoptosisrelated factors including Bcl-XL and Caspase-3, Alzheimer's disease pathological biomarkers including Aβ1-42, p-Tau and its downstream NF-κB/TNF-α signalling pathway. Results: Sch B can promote the depolymerization of Aβ oligomers, increase the cell proliferation rates of SH-SY5 Y induced by Aβ, inhibit apoptosis of SH-SY5 Y after injuried by Aβ, it can increase expression of Bcl-XL and reduce expression of Caspase-3, inhibit expression of Aβ1-42 and p-Tau, down-regulated expression of NF-κB/TNF-α signaling pathway. Conclusion: Sch B could target the affinity of Aβ ligands and promote the depolymerization of Aβ oligomers, and inhibit the apoptosis of neurons. The mechanism might be related to the depolymerization and downregulation of oligomers neuron Aβ expression, inhibition of Tau hyperphosphorylation, and further down-regulation of NF-κB/TNF-α pathway.

关 键 词：五味子乙素 Β-淀粉样蛋白 NF-κB/TNF-α信号通路 凋亡 受损神经元 

分 类 号：R285[医药卫生—中药学]