基于网络药理学、分子对接及实验验证探讨刺梨黄酮干预溃疡性结肠炎的作用机制

Exploring the Action Mechanism of Rosa roxburghii Fruit Flavonoids in the Intervention of Ulcerative Colitis Based on Network Pharmacology,Molecular Docking and Experimental Verification

运用网络药理学方法和分子对接技术探讨刺梨黄酮干预溃疡性结肠炎(ulcerative colitis,UC)的核心成分和作用机制。用乙醇-纤维素酶法提取刺梨黄酮,并采用液相色谱-质谱法鉴定刺梨黄酮提取物主要的化学成分,然后通过分析平台筛选得到刺梨黄酮中34个活性成分和146个干预溃疡性结肠炎作用的靶点,构建“刺梨黄酮-活性成分-交集靶点-溃疡性结肠炎”网络。基因本体通路分析和京都基因与基因组百科全书通路富集分析结果显示,刺梨黄酮干预UC主要涉及花生四烯酸代谢通路、核因子κB信号通路、癌症通路等信号通路;对网络筛选的主要活性成分和关键靶点做分子对接,对接能均小于-6.0kcal/mol,显示刺梨黄酮筛选的核心活性成分和治疗UC的关键靶点有较强的亲和能力,说明网络分析结果可靠。动物实验结果显示刺梨黄酮干预可有效缓解小鼠体质量下降与结肠组织病变,并可显著抑制模型组炎症因子肿瘤坏死因子α、白细胞介素-1β和白细胞介素-6的表达(P<0.05),且Westernblot结果显示刺梨黄酮可有效地抑制花生四烯酸代谢通路的关键靶点蛋白环氧合酶-2和5-脂氧合酶的表达。综上,刺梨黄酮通过多成分-多靶点-多通路协同作用干预UC,研究结果可为刺梨功能性食品的开发与利用提供理论依据。

The aim of this study was to investigate the core components and mechanism of action of Rosa roxburghii fruit flavonoids in the intervention of ulcerative colitis(UC)using network pharmacology and molecular docking.We extracted flavonoids from R.roxburghii fruit by an ethanol-cellulase method and used liquid chromatography-mass spectrometry(LC-MS)to identify the major chemical components of the flavonoid extract,and then we selected 34 active flavonoid components and 146 targets in the intervention of UC on an analytical platform and constructed a“R.roxburghii flavonoids-active components-intersecting targets-UC”network.The results of Gene Ontology pathway analysis and Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway enrichment analysis showed that the intervention of R.roxburghii flavonoids in UC mainly involved the arachidonic acid signaling pathway,the nuclear factor-kappa B signaling pathway,and cancer pathways.Molecular docking was performed on the major active ingredients and the key targets selected from the network,with docking energies all less than-6.0 kcal/mol,indicating a strong binding affinity between the core active ingredients and the key targets for UC treatment.So,the network analysis results are reliable.The results of animal experiments showed that intervention with R.roxburghii flavonoids could effectively alleviate the body mass loss and colon pathological changes of UC mice,and significantly inhibit the expression of inflammatory cytokines such as tumour necrosis factor-α,interleukin(IL)-1βand IL-6 in the UC model group(P<0.05).The results of Western blot showed that R.roxburghii flavonoids could effectively inhibit the expression of the key target proteins associated with the metabolic pathway of arachidonic acid,such as cyclooxygenase-2 and 5-lipoxygenase.In conclusion,R.roxburghii flavonoids can intervene in UC through a multi-component,multi-target and multi-pathway mechanism.The results of this research can provide a theoretical basis for the development and utilization of functional foods from R.roxburghii.