基于网络药理学探讨青钱柳调节血糖的作用机制＊

目的 通过网络药理学及分子对接的方法探讨青钱柳调节血糖的潜在作用机制。方法 通过文献对青钱柳进行化学成分搜集,利用Swiss ADME平台进行活性成分筛选,借助Swiss Target Prediction数据库进行作用靶点进预测,运用Cytoscape 3.8构建药物成分-靶点网络;检索Genecards,OMIM以及Disgenet数据库获取血糖靶点;通过韦恩图,得到青钱柳调节血糖靶点;使用String平台构建蛋白质-蛋白质相互作用（PPI）网络;使用metascape数据平台进行基因分类（GO）富集分析和京都基因和基因组百科全书（KEGG）通路分析。构建青钱柳调节血糖活性成分-共同靶点-信号通路网络。同时进行分子对接验证。建立实验性Ⅱ型糖尿病大鼠模型,检测大鼠空腹血糖（FBG）、空腹胰岛素（FIns）、糖化血红蛋白（HbA1c）水平,及大鼠胰脏组织ESR2、PTPN1的表达。结果 通过文献搜集筛选获得青钱柳39个活性成分,导入Swiss Target Prediction平台获得509个相关靶点。检索数据库获得血糖相关靶点1618个。二者取交集即为青钱柳调节血糖作用靶点134个,经PPI结果筛选得治疗靶点24个。GO富集分析共得主要条目中生物过程20条,细胞组成7条,分子功能10条。KEGG 通路分析共得到核心信号通路8条。将8条通路涉及靶点与药物成分-靶点网络merge取交集后可得核心靶点18个,靶点连接的核心治疗成分29个。分子对接结果表明山奈酚、咖啡酸、槲皮素等核心成分与血糖调节相关蛋白及作用靶点具有较强的结合活性,可能具备相应的调节效果。青钱柳多糖能明显降低实验性Ⅱ型糖尿病大鼠模型FBG、FIns、HbA1c水平,提高大鼠胰脏组织ESR2、PTPN1的表达。结论 青钱柳可能是通过调节雌激素受体2型（ESR2）、酪氨酸蛋白磷激酶1型（PTPN1）、基质金属蛋白酶2型（MMP2）、金属蛋白酶2型（MMP9）、糖酶合酶激酶3B（GSK3B）、胰岛素样生长因子1受体（IGF1R）等靶点,在体内参与神经信号传导,发挥相关蛋白质合成,细胞间信号传递等功能来调节血糖。

Mechanism of Cyclocarya Paliurus Regulating Blood Glucose Based on Network Pharmacology

Objective To explore the potential mechanism of Cyclocarya Paliurus in regulating blood glucose by network pharmacology and molecular docking.Methods The chemical constituents of Cyclocarya Paliurus were collected and active components were screened by Swiss ADME platform. The target prediction database of Swiss was used to predict the target, and Cytoscape 3.8 was used to construct the drug- target network; Genecards, OMIM and digenet databases were helping to find blood glucose targets; to obtain the blood glucose regulating targets of Cyclocarya Paliurus through Wayne map; The protein- protein interaction (PPI) network was constructed by using string platform; The gene ontology (GO) analysis and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis were performed by using metascape data platform, which construct a common target signal pathway network of active components regulating blood glucose in Cyclocarya Paliurus. At the same time molecular docking verification was carried out. An experimental type 2 diabetic rat model was established. Fasting blood glucose (FBG), fasting insulin (FIns), glycosylated hemoglobin (HbA1c) levels and expression of ESR2 and PTPN1 in pancreas tissue of rats were detected.Results Through literature collection and screening, 39 active ingredients of Cyclocarya Paliurus were obtained, and 509 related targets were obtained by importing them into the Swiss Target Prediction platform. The database was searched to obtain 1,618 blood glucose-related targets. The intersection of the two was the 134 targets of Cyclocarya Paliurus for regulating blood sugar, and 24 therapeutic targets are screened by the PPI results. In the GO enrichment analysis, there were 20 biological processes, 7 cell composition, and 10 molecular functions in the main items. A total of 8 core signal pathways were obtained from KEGG pathway analysis. The 8 pathways involving targets and the drug component-target network merge can get 18 core targets and 29 core therapeutic components connected to the targets. The results of molecular docking showed that core components such as kaempferol, caffeic acid, and quercetin had strong binding activity with blood glucose regulation-related proteins and targets, which may have corresponding regulatory effects. Polysaccharides from P. salicis significantly decreased the levels of FBG, FIns and HbA1c in experimental type 2 diabetic rats, and increased the expression of ESR2 and PTPN1 in pancreas tissue of rats.Conclusion Cyclocarya Paliurus may regulate blood glucose by regulating Estrogen receptor beta (ESR2), Tyrosine-protein phosphatase non-receptor type 1 (PTPN1), Matrix metalloproteinase-2 (MMP2), Matrix metalloproteinase-9 (MMP9), Glycogen synthase kinase-3 beta (GSK3B), Insulin-like growth factor 1 receptor (IGF1R) and other targets, which participate in nerve signal transduction in vivo, play related protein synthesis, intercellular signal transmission to regulate blood glucose.