基于网络药理学和分子对接预测何首乌改善脂质代谢紊乱作用机制及调脂作用研究

The mechanism of Polygonum multiflorum Thunb.to attenuate lipid metabolism disorder based on network pharmacology

目的:应用网络药理学和分子对接方法预测何首乌改善脂质代谢紊乱的作用机制。方法:借助软件ChemDraw绘制何首乌化学成分,基于Swiss ADME,Swiss Target Prediction平台与Genecards, DisGeNET数据库筛选出活性成分与改善脂质代谢紊乱的潜在靶点,通过软件Cytoscape和STRING数据库分别构建“药物-活性成分-潜在靶点”网络结构模型与蛋白质互作网络(PPI)网络,运用DAVID数据库对潜在靶点进行基因本体论(GO)与京都基因与基因组百科全书(KEGG)富集分析。借助软件PyMOL,AutoDock进行活性成分与核心靶点分子对接。长期灌胃给药β-淀粉样前体蛋白/早老蛋白-1双转基因小鼠(APP/PS1小鼠)并统计其体质量和体脂率,检测血脂[三酰甘油(TG)、总胆固醇(TC)、高密度脂蛋白胆固醇(HDL-C)、低密度脂蛋白胆固醇(LDL-C)]水平。结果:筛选出30种何首乌活性成分以及143个治疗脂质代谢紊乱潜在靶点,富集分析显示何首乌与脂质代谢紊乱的共同生物学过程与蛋白磷酸化、蛋白结合、酶结合与蛋白激酶活性等相关,作用的主要通路为磷脂酰肌醇3-激酶-蛋白激酶(PI3K-Akt)、缺氧诱导因子(HIF)-1、雌激素(estrogen)信号通路等。分子对接预测活性成分与核心靶点间稳定连接结构关系。动物实验结果显示,与模型组相比,各药物组体重无显著差异,但体脂率显著降低(P<0.05);蒸晒品对TG,LDL-C异常升高具有显著降低作用,但生品仅对TG有影响(P<0.05)。结论:何首乌有确切降脂作用,预测其机制为ω-羟基大黄素-8-甲醚、苜蓿素、山奈酚等3种主要活性成分通过与表皮生长因子受体(EGFR)、雌激素受体1(ESR1)、基质金属蛋白酶9(MMP9)等3个核心靶点结合,影响PI3K-Akt, HIF-1,estrogen信号通路来发挥。

Objective:To predict the mechanism of Polygonum multiflorum Thunb.on attenuating lipid metabolism disorder by applying Network Pharmacology and Molecular Docking.Methods:Using software ChemDraw,the chemical constituents of Polygonum multiflorum Thunb.were drawn and documented.Main active ingredients and potential targets of attenuating lipid metabolism disorder were screened out based on the databases like Swiss ADME,Swiss Target Prediction platform and Genecards,and DisGeNET.The drug-active ingredients-potential targets network structure model and PPI network are constructed through the software Cytoscape and STRING database,respectively.Potential targets were analyzed by GO enrichment analysis,KEGG pathway enrichment analysis and visualized.Using software PyMOL and AutoDock,the active ingredients were docked with core target molecules.The body weight and body fat rate of APP/PS1 mice with long-term intragastric administration were measured and the content of serum lipid(TG,TC,HDL-C,LDL-C)were detected.Results:A total of 30 active ingredients of Polygonum multiflorum Thunb.and 143 potential targets for the treatment of lipid metabolism disorder were obtained.The enrichment analysis shows that the common biological process of Polygonum multiflorum Thunb.-and metabolic disorders are related to protein phosphorylation,protein binding,enzyme binding,protein kinase activity biological processes,and the signalling pathways of PI3K-Akt,HIF-1,estrogen are mainly involved in the major therapeutic role.Molecular docking predicts the stable connection structure between the active component and the core targets.The results of animal experiments showed that compared with the model group,there was no significant difference in weight between drug administered groups and the body fat rate decreased significantly(P<0.05).Steaming and sun-buring products had significant effect on the abnormal increase of TG and LDL-C,but crude Polygonum multiflorum Thunb.only had an impact on TG(P<0.05).Conclusion:Polygonum multiflorum Thunb.has an significant lipid-attenuating effect.The mechanism is predicted to be related with three main active ingredients,including omega-hydroxytroxanin-8-methyl ether,Tricin and Kaempferolare,and PI3K-Akt,HIF-1 and Estrogen signaling pathways are affected through three core targets of ECFR,ESR1 and MMP9.