基于模式生物斑马鱼研究姜黄抗血管新生的作用及机制

Effect and mechanism of Curcuma longa anti-angiogenesis based on model biology zebrafish

目的利用模式生物斑马鱼研究姜黄Curcuma longa抗血管新生作用,并采用网络药理学和分子对接技术探究姜黄抗血管新生的作用机制。方法采用模式生物斑马鱼模型,考察姜黄挥发油、姜黄素以及姜黄水提液对斑马鱼节间血管生长的抑制作用,检测各给药组斑马鱼节间血管生成数,计算血管生成抑制率。利用网络药理学和分子对接技术预测姜黄抗血管新生的作用机制,构建"药物-成分-靶点"网络和蛋白质-蛋白质相互作用(protein-protein interaction,PPI)网络,进行基因本体(gene ontology,GO)富集分析及京都基因与基因组百科全书(Kyoto encyclopedia of genes and genomes,KEGG)通路富集分析;通过AutoDock vina软件对活性成分与关键靶点进行分子对接验证。结果姜黄挥发油、姜黄素以及姜黄水提液均可明显抑制斑马鱼血管新生(P<0.05),呈剂量相关性,其中姜黄素抑制血管生成作用最强。网络药理学预测发现姜黄共有21种主要活性成分,可通过调节前列腺素G/H合酶2(prostaglandin G/H synthase 2,PTGS2)、维生素D3受体(vitamin D3 receptor,VDR)、丝氨酸/苏氨酸蛋白激酶(serine/threonine kinase 1,AKT1)和血清白蛋白(albumin,ALB)等关键靶点,进而调控磷脂酰肌醇-3-激酶(phosphatidylinositide 3-kinases,PI3K)-AKT和血管内皮生长因子(vascular endothelial growth factor,VEGF)等相关信号通路,从而发挥抑制血管新生的作用。分子对接结果显示姜黄的主要活性成分与核心靶点均能自发结合。结论姜黄挥发油、姜黄素以及姜黄水提液对斑马鱼血管新生的抑制作用存在差异,姜黄抗血管新生具有多成分、多靶点、多途径的特点,为姜黄对血管新生依赖性疾病的防治提供了参考。

Objective To study the anti-angiogenesis effect of Jianghuang(Curcuma longa)by model biology zebrafish,and predict the mechanism based on network pharmacology and molecular docking technology.Methods The inhibitory effect of volatile oil of C.longa,curcumin and decoction of C.longa on intersegmental vascular growth of zebrafish were studied by model biology zebrafish.The number of intact vessels was counted and inhibitory rate of angiogenesis was calculated.Network pharmacology and molecular dockingtechnology were used to predict the anti-angiogenesis mechanism of C.longa."Drug-component-target"and protein-protein interaction(PPI)network were constructed.Gene ontology(GO)enrichment analysis and Kyoto encyclopedia of genes and genomes(KEGG)pathway enrichment analysis were performed.AutoDock vina was used to verify the molecular docking between active ingredients and key target.Results The volatile oil of C.longa,curcumin and decoction of C.longa significantly inhibited zebrafish angiogenesis with dose-dependent(P<0.05),and curcumin had the strongest inhibitory effect on angiogenesis.Network pharmacology predicted that there were 21 main active ingredients in C.longa,which regulated phosphatidylinositide 3-kinases-serine/threonine kinase(PI3 K-AKT),vascular endothelial growth factor(VEGF)and other related signal pathways by regulating key targets such as prostaglandin G/H synthase 2(PTGS2),vitamin D3 receptor(VDR),AKT1,and albumin(ALB),thereby inhibited angiogenesis.Molecular docking showed that main active ingredients and key targets could be spontaneously combined..Conclusion The volatile oil of C.longa,curcumin and decoction of C.longa had different inhibitory effects on angiogenesis of zebrafish,anti-angiogenesis effect of C.longa has characteristics of multiple components,multiple targets and multiple pathways,which provides a reference for prevention and treatment of angiogenesis dependent diseases.