基于网络药理学和分子对接技术探究槐花散治疗溃疡性结肠炎的作用机制＊

目的 该研究采用网络药理学和分子对接的研究方法结合体内实验探讨槐花散治疗溃疡性结肠炎（ulcerative colitis, UC）的主要化合物和靶点,并分析其作用机制。方法 利用多个数据库筛选药物活性成分、药物靶点和UC疾病基因,并进行拓扑分析,构建“化合物-药物靶点-疾病基因”关系网络。利用DAVID在线分析工具对候选靶点进行基因本体（Gene ontology, GO）功能和生物通路（KEGG）富集分析。通过SYBYL X2.0软件进行分子对接,获取总评分、共识评分和结合位点。利用硫酸葡聚糖钠盐（dextran sodium sulfate,DSS）建立UC小鼠模型,槐花散干预后,评估疾病活动指数和结肠病理,RT-qPCR检测白介素6（IL-6）、白介素17（IL-17）mRNA表达水平,Western Blot检测IL-6、IL-17蛋白表达水平。结果 槐花散中有20个化合物对应375个靶点对121个UC疾病基因产生影响。对72个关键靶点作GO功能分析筛选得303个GO条目,其中生物过程相关的条目223条,细胞组成相关的条目25条,分子功能相关的条目55条。KEGG富集分析筛选得到88条通路。分子对接结果显示关键靶点与对应的化合物有较好的结合活性。体内实验显示,槐花散可以改善UC小鼠DAI评分,下调IL-6、IL-17的表达,减轻结肠组织病理损伤。结论 槐花散治疗UC具有多成分、多靶点、多通路的特点,为槐花散治疗UC的进一步研究提供了理论依据。

Explore Mechanism of Huaihua Powder for Treatment of Ulcerative Colitis Based on Network Pharmacology and Molecular Docking

Objective To explore the main compounds and targets of Huaihua Powder and analyzed its mechanism in the treatment of ulcerative colitis (UC) based on network pharmacology, molecular docking and in vivo experiment.Methods The active pharmaceutical ingredients, drug targets and UC disease genes were selected from multiple databases. All of these were topologically analyzed, constructing a compound-target-gene network. Gene ontology (GO) and biological pathway (KEGG) enrichment analysis of candidate targets were carried out by the DAVID online analysis tool. SYBYL X2.0 software was used for molecular docking to obtain Total Score, CScore and binding sites. Dextran sodium sulfate (DSS) was used to establish the mouse UC models. After the intervention of Huaihua Powder, the disease activity index and colon pathology were evaluated. RT-qPCR was used to detect the mRNA expression of interleukin-6 (IL-6) and interleukin-17 (IL-17). Western Blot was used to detect the protein expression of IL-6 and IL-17.Results In Huaihua Powder, 375 targets which corresponding to 20 compounds affected 121 disease genes of UC. A total of 303 gene ontology terms of 72 key-targets were screened out, including 223 biological process terms, 25 cellular component terms, and 55 molecular function terms. We screened 88 pathways by KEGG enrichment analysis of candidate genes. Molecular docking results showed that the key targets had better binding activity with the corresponding compounds. In vivo experiments showed that Huaihua Powder could improve the DAI score of UC mice, down-regulate the expression of IL-6 and IL-17 and reduce the pathological damage of colon tissue.Conclusion Huaihua Powder has the characteristics of multi-components, multi-targets and multi-pathways in the treatment of UC, which provides a theoretical basis for further research on the treatment of UC by Huaihua Powder.