基于网络药理学及分子对接探讨黄连治疗2型糖尿病的作用机制

Mechanism of Coptis chinensis in Treatment of Type 2 Diabetes Mellitus Based on Network Pharmacology and Molecular Docking

目的:通过网络药理学及分子对接探讨黄连治疗2型糖尿病的作用机制。方法:应用中药系统药理学数据库与分析平台(TCMSP),设定药物口服生物利用度(OB)≥30%和药物相似性(DL)≥0.18筛选出黄连的活性化学成分,利用UniProt数据库和CTD数据库,检索出活性成分对应的作用靶点以及2型糖尿病的相关靶点;利用Cytoscape 3.6.1软件制作“黄连-活性成分-潜在靶点”可视化网络图,结合STRING数据库构建交集靶点的PPI网络图,对核心靶点进行GO功能富集和KEGG通路富集分析。另对核心靶基因与黄连的活性成分进行分子对接,验证活性成分与核心靶基因的结合能力。结果:①筛选出黄连的活性成分共有14种,治疗对应的2型糖尿病靶点189个,其中潜在靶点136个,包括核心靶点20个(degree≥12),有显著意义的信号通路137条。②基因本体(GO)功能富集分析显示,黄连在治疗2型糖尿病的预测靶点所参与的生物学过程(BP)方面涉及腺发育、上皮细胞增殖、DNA结合转录因子活性的调节、氧化应激及对脂多糖等多种细胞凋亡反应;在分子功能(MF)方面涉及DNA转录因子结合、酶结合、受体结合等反应;在细胞组成(CC)方面涉及酶调节复合物、囊腔、膜微区、膜区的构成。③“成分-靶点-通路”网络分析发现,黄连治疗2型糖尿病的主要潜在活性成分可能为槲皮素、氢化小檗碱、小檗碱等,其可作用于丝裂原活化蛋白激酶1(MAPK1)、丝氨酸/苏氨酸蛋白激酶1(AKT1)、肿瘤抗原p53(TP53)、转录因子AP-1(JUN)、肿瘤坏死因子(TNF)、白介素-6(IL-6)等核心靶点,从而对MAPK、糖基化终末产物(AGEs)-RAGE、PI3K-Akt、Toll样受体、HIF-1、TNF等信号通路进行调节,起到治疗2型糖尿病的作用。④分子对接结果显示,黄连的活性成分与上述核心靶点的分子对接亲和力均<-7 kcal/mol(1 kca/mol=4.186 kJ/mol),结合活性较好。结论:黄连可能通过调控糖脂代谢、调控炎症反应及细胞增殖与凋亡、促进氧化应激等多方面以发挥治疗2型糖尿病及其并发症的作用。

Objective: To explore the mechanism of Coptis chinensis in the treatment of type 2 diabetes mellitus based on network pharmacology and molecular docking. Methods: The active chemical ingredients of Coptis chinensis were screened through the Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform(TCMSP), oral bioavailability(OB) ≥30%and drug-likeness(DL) ≥0.18 were set. The corresponding target of active ingredients and related targets of type 2 diabetes mellitus were retrieved through UniProt database and CTD database;Cytoscape 3.6.1 software was used to make the visual network diagram of "Coptis chinensis-active ingredients-targets", and STRING database was used to construct the PPI network diagram of intersection targets, and the core targets were analyzed by GO function enrichment and KEGG pathway enrichment. In addition, molecular docking was conducted between the core targets and the active ingredients of Coptis chinensis to verify the binding ability between the active ingredients and the core targets. Results:(1) A total of 14 active ingredients of Coptis chinensis, 189 targets for the treatment of type 2 diabetes mellitus were screened out, among which 136 potential targets were identified, including 20 core targets(degree≥12), 137 signaling pathways with significant significance.(2) Gene Ontology(GO) functional enrichment analysis showed that,the biological processes(BP) involved in the predicted target of Coptis chinensis in the treatment of type 2 diabetes mellitus involved gland development, epithelial cell proliferation, regulation of DNA-binding transcription factor activity, oxidative stress and apoptosis response to lipopolysaccharide, etc. In terms of molecular function(MF), it involved DNA transcription factor binding, enzyme binding, receptor binding and other reactions. In terms of cell components(CC), it involved the composition of enzyme regulated complex, cystic cavity, membrane microregion and membrane region.(3) Components-target-pathway network analysis showed that, the main potential active ingredients of Coptis chinensis in the treatment of type 2 diabetes mellitus were quercetin, berberine,etc. It can act on mitogen-activated proteinase 1(MAPK1), serine/threonine protein kinase(AKT1), tumor protein p53(TP53), transcription factor AP-1(JUN), tumor necrosis factor(TNF), interleukin-6(IL-6), which can regulate MAPK, AGEs-RAGE, PI3 K-Akt,Toll-like receptors, HIF-1, TNF signaling pathways, etc.(4) The molecular docking results showed that, the molecular docking affinity between the active ingredients of Coptis chinensis and the above core targets was less than-7.0 kcal/mol(1 kca/mol=4.186 kJ/mol), and the binding activity was better. Conclusion: Coptis chinensis may play an important role in the treatment of type 2 diabetes mellitus and its complications by regulating glucose and lipid metabolism, inflammatory response, cell proliferation and apoptosis and promoting oxidative stress, etc.