基于网络药理学和体外实验探究芍药内酯苷改善阿尔茨海默病的作用机制

Mechanism of albiflorin in improvement of Alzheimer′s disease based on network pharmacology and in vitro experiments

基于网络药理学、分子对接技术和体外实验研究芍药内酯苷治疗阿尔茨海默病(Alzheimer′s disease,AD)的作用机制。采用网络药理学预测芍药内酯苷抗AD的潜在作用靶点和通路,结合分子对接技术对芍药内酯苷与关键靶蛋白进行分子对接验证,最后使用芍药内酯苷干预Aβ25-35诱导大鼠嗜铬瘤细胞(PC12细胞)所构建的AD细胞模型,以验证核心靶点和通路。网络药理学分析结果显示,芍药内酯苷主要作用于丝裂原活化蛋白激酶1(MAPK1,又称ERK2)、白蛋白(ALB)、表皮生长因子受体(EGFR)、半胱氨酸天冬氨酸蛋白酶3(CASP3)、5-羟色胺转运体(SLC6A4)等关键靶点和MAPK、cAMP、cGMP-PKG等信号通路。分子对接结果表明芍药内酯苷与MAPK1(ERK2)有较好的结合性。体外实验显示,与空白组相比较,模型组细胞活力显著下降,B淋巴细胞瘤-2(Bcl-2)表达水平显著降低,Bcl-2关联X蛋白(Bax)表达水平显著升高,细胞外信号调节蛋白激酶1/2(ERK1/2)磷酸化水平和p-ERK1/2与ERK1/2相对表达量比值均显著下降;与模型组相比较,芍药内酯苷组细胞活力显著升高,Bcl-2的表达量上调,Bax表达量下调,ERK1/2磷酸化水平和p-ERK1/2与ERK1/2相对表达量比值显著增加。研究结果提示芍药内酯苷抗AD的作用机制可能与其激活MAPK/ERK信号通路,进而抑制神经细胞凋亡有关。

This study aimed to explore the mechanism of albiflorin in the treatment of Alzheimer′s disease(AD)based on network pharmacology,molecular docking,and in vitro experiments.Network pharmacology was used to predict the potential targets and pathways of albiflorin against AD,and molecular docking technology was used to verify the binding affinity of albiflorin to key target proteins.Finally,the AD cell model was induced by Aβ_(25-35)in rat pheochromocytoma(PC12)cells and intervened by albiflorin to validate core targets and pathways.The results of network pharmacological analysis showed that albiflorin acted on key targets such as mitogen-activated protein kinase-1(MAPK1 or ERK2),albumin(ALB),epidermal growth factor receptor(EGFR),caspase-3(CASP3),and sodium-dependent serotonin transporter(SLC6A4),and signaling pathways such as MAPK,cAMP,and cGMP-PKG.The results of molecular docking showed that albiflorin had strong binding affinity to MAPK1(ERK2).In vitro experiments showed that compared with the blank group,the model group showed decreased cell viability,decreased expression level of B-cell lymphoma 2(Bcl-2),increased Bcl-2-associated X protein(Bax),and reduced phosphorylation level of extracellular signal-regulated kinase 1/2(ERK1/2)and the relative expression ratio of p-ERK1/2 to ERK1/2.Compared with the model group,the albiflorin group showed potentiated cell viability,up-regulated expression of Bcl-2,down-regulated Bax,and increased phosphorylation level of ERK1/2 and the relative expression ratio of p-ERK1/2 to ERK1/2.These results suggest that the mechanism of albiflorin against AD may be related to its activation of the MAPK/ERK signaling pathway and its inhibition of neuronal apoptosis.