基于网络药理学及分子对接探讨血府逐瘀汤加减治疗糖尿病周围神经病变的机制

Exploration on Mechanism of Modified Xuefu Zhuyu Decoction in Treatment of Diabetic Peripheral Neuropathy Based on Network Pharmacology and Molecular Docking

目的:通过网络药理学的方法,对血府逐瘀汤加减治疗糖尿病周围神经病变的机制进行探索。方法:利用中药系统药理学技术平台(Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform,TCMSP)数据库对血府逐瘀汤加减的组方药物进行查询,筛选其化学活性物质和靶标基因,并利用基因组注释数据库平台(Genecards)预测疾病的靶点基因,药物与疾病的靶标基因取交集后构建活性成分-靶点网络图;构建PPI网络,并寻找核心基因,再进行GO富集分析和KEGG富集分析,找出所涉及的信号通路,构建靶点-通路网络图,利用分子对接技术验证小分子和蛋白的对接情况,最后进行动物实验验证血府逐瘀汤加减对糖尿病周围神经病变(Diabetic peripheral neuropathy,DPN)大鼠蛋白表达的影响。结果:血府逐瘀汤加减治疗DPN的关键化学成分共80个,分别为槲皮素、木犀草素、三柰酚、汉黄芩素、黄芩素等;药物-DPN的共同靶点142个,关键蛋白14个。GO富集结果包括小梁形态发生、积极调控、膜筏等;KEGG富集结果包括AGE-RAGE信号通路、流体剪切应力与动脉粥样硬化、IL-17信号通路等。分子对接显示槲皮素、木犀草素、三柰酚和汉黄芩素与AKT1结合活性较强;槲皮素、甘草查尔酮A与RELA结合活性较强;甘草查尔酮A与MAPK14、ESR1结合活性较强。与模型对照组相比,血府逐瘀汤17.1 g/kg组能明显降低大鼠机械痛阈值,下调AKT1、RELA、MAPK14的蛋白相对表达(P<0.05)。结论:血府逐瘀汤加减可能通过槲皮素、木犀草素、三柰酚等多种化学成分调节AKT1、RELA、MAPK14等靶点,进而影响多条信号通路,通过抗炎、促进神经细胞增殖,修复神经组织共同发挥着治疗DPN的作用。

Objective:To explore the mechanism of modified Xuefu Zhuyu Decoction in the treatment of diabetic peripheral neuropathy(DPN)based on network pharmacology.Methods:The components of modified Xuefu Zhuyu Decoction were searched from the Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform(TCMSP),followed by the screening of its chemical active substances and target genes.The DPN-related target genes were retrieved from Genecards.The modified Xuefu Zhuyu Decoction and DPN target genes were intersected to construct the active ingredient-target network.PPI network was constructed,and the core genes were selected and subjected to GO and KEGG enrichment analysis to find out the related signaling pathways.After the target-pathway network was constructed,the molecular docking technology was used to verify the docking of small molecules to proteins.The effects of modified Xuefu Zhuyu Decoction on protein expression in DPN rats were verified by animal experiments.Results:A total of 80 key chemical components for the treatment of DPN by modified Xuefu Zhuyu Decoction were observed,including quercetin,luteolin,kaempferol,wogonin,and baicalein.There were 142 DPN-related common targets and 14 key proteins.The GO enrichment analysis revealed trabecular morphogenesis,active regulation,membrane rafts,etc.As demonstrated by KEGG enrichment analysis,the AGE-RAGE signaling pathway,fluid shear stress and atherosclerosis,and IL-17 signaling pathway were involved.Molecular docking showed the strong binding activities of quercetin,luteolin,kaempferol,and wogonin to AKT1,of quercetin and licochalcone A to RELA,and of licochalcone A to MAPK14 and ESR1.Compared with the model group,the modified Xuefu Zhuyu Decoction at 17.1 g/kg significantly decreased the mechanical pain threshold of rats and down-regulated the relative protein expression levels of AKT1,RELA,and MAPK14(P<0.05).Conclusion:Modified Xuefu Zhuyu Decoction may regulate such targets as AKT1,RELA,and MAPK14 through various chemical components like quercetin,luteolin,and kaempferol to affect multiple signaling pathways,thus inhibiting inflammation,promoting nerve cell proliferation,repairing nerve tissue,and alleviating DPN.