结合网络药理学及动物实验初步探究苍术抗溃疡性结肠炎的作用机制

Elucidating the therapeutic mechanism of Atractylodes lancea for ulcerative colitis based upon network pharmacology and animal experiments

目的:基于网络药理学方法结合体内动物试验探究苍术抗溃疡性结肠炎的作用机制。方法:(1)通过TCMSP、BATMAN-TCM数据库筛选出苍术的活性成分及其可能的作用靶点;通过GeneCards数据库检索溃疡性结肠炎(ulcerative colitis,UC)疾病的相关靶点;将上述靶点导入Venny 2.1中对苍术活性成分的作用靶点与UC的疾病靶点取交集,获得苍术抗UC的交集靶点;采用Cytoscape 3.7.1构建苍术抗UC的“苍术-活性成分-交集靶点”网络,分析其关键活性成分;将交集靶点导入STRING数据库,构建蛋白-蛋白互作网络图,筛选出核心靶点;将交集靶点导入Matescape数据库对其进行GO功能及KEGG富集分析,最后使用Maestro 11.1软件对关键活性成分与核心靶点蛋白进行分子对接。(2)体内复制葡聚糖硫酸钠(dextran sodium sulfate,DSS)诱导UC小鼠模型,将BALB/c小鼠通过随机数字表法分为空白组、模型组、苍术醇提物组(1110 mg·kg^(-1)·d^(-1))及阳性药组(柳氮磺吡啶,SASP,250 mg·kg^(–1)·d^(–1)),每组6只;灌胃给药(10 mL·kg^(-1)),每天1次,连续给药7 d,末次给药1 h后处死,并进行相关指标检测。结果:(1)苍术抗UC的活性成分有26个,苍术-UC交集靶点273个;KEGG富集分析显示主要涉及的信号通路有MAPK通路及PI3K/Akt等炎症通路;分子对接结果显示其关键活性成分与核心靶点蛋白之间具有较强的结合活性。(2)动物实验数据表明,苍术醇提物可明显增加UC小鼠的结肠长度、降低疾病活动指数评分,减缓UC小鼠结肠组织病变程度、增加杯状细胞的个数,抑制结肠组织中IL-1β、MMP-2、MMP-9等炎性介质的高表达,调控PI3K/Akt通路的激活。结论:该研究结合网络药理学及动物实验初步证实苍术可能通过作用于IL-1β等靶点、参与PI3K/Akt信号通路的调控改善UC小鼠结肠损伤,为深入研究苍术及其活性成分抗溃疡性结肠炎的作用机制提供实验依据和理论参考。

OBJECTIVE To elucidate the therapeutic mechanism of Atractylodes lancea for ulcerative colitis(UC)based upon network pharmacology and in vivo experiments.METHODS The active ingredients and potential targets of Atractylodes lancea were selected through the databases of TCMSP and BATMAN-TCM.And the relevant targets of UC were retrieved through the databases of OMIM and GeneCards.Then the above targets were entered into Venny 2.1 for intersecting with important disease targets of UC.For obtaining a common target of Atractylodes lancea against UC,a“Atractylodes lancea-active ingredient-common target”network of Atractylodes lancea against UC was constructed with Cytoscape 3.7.1 and its key active ingredients were examined.Common targets were imported into the database of STRING for constructing a protein-protein interaction(PPI)network diagram and screening out core targets.Common targets were imported into the database of Matescape for Gene Ontology(GO)function and Kyoto Encyclopedia of Genes and Genomes(KEGG)enrichment analysis.Then Maestro 11.1 software was utilized for molecular docking between the active components of Atractylodes lancea and the core targets.After an induction of dextran sodium sulfate(DSS),BALB/c mice were randomized into four groups of control group,model,Atractylodes lancea extract(1110 mg·kg^(-1)·d^(-1))and sulfasalazine(SASP,250 mg·kg^(-1)·d^(-1))(n=6 each).After a once daily gavage(10 mL·kg^(-1))for 7 consecutive days,the animals were sacrificed at Hour 1 after the last dosing with relevant indicators tested.RESULTS There were 26 potential active ingredients of Atractylodes lancea against UC and 273 drug disease common targets.KEGG enrichment analysis revealed that the major signaling pathways of cancer,MAPK and PI3K/Akt.The result of molecular docking indicated a strong affinity between its key components and core targets.Animal experiments showed that Atractylodes lancea could significantly enhance colonic length,lower DAI scores and lessen the degree of colonic tissue lesions,boost the number of goblet cells,suppress the high expressions of IL-1β,MMP-2 and MMP-9 in colonic tissue and modulate the activation of PI3K/Akt signaling pathway.CONCLUSION This study has preliminarily confirmed that Atractylodes lancea could improve colonic injury in UC through participating in the regulation of PI3K/Akt signaling pathway and inflammatory targets(e.g.IL-1β).It provided scientific and theoretical rationales for in-depth researches on the mechanism of Atractylodes lancea and its active ingredients against UC.