目的 运用网络药理学和实验验证的方法分析马齿苋治疗结直肠癌(Colorectal cancer，CRC)的潜在作用机制。方法 借助TCMSP、ETCM数据库获取马齿苋的有效成分及靶点，通过Genecards、Drugbank、OMIM数据库检索与CRC相关的靶点，将两者的靶点导入Venny数据库、String数据库和Cytoscape平台构建PPI蛋白互作网路，用Metascape数据库和微生信平台对共同核心靶点进行GO功能和KEGG通路富集分析，并将有效成分和关键靶点使用Autodock和Pymol软件进行分子对接。构建AOM/DSS诱导的小鼠CRC模型，给予马齿苋干预，观察其结肠组织病理变化，使用qRT-PCR实验、Western blot实验对关键靶点进行验证。此外，采用槲皮素、木犀草素、山柰酚作用于HCT116细胞，观察对细胞增殖影响并计算IC50，并使用qRT-PCR实验、Western blot实验进行验证。结果 网络药理学获得马齿苋的有效成分11个和对应靶点209个靶点，疾病靶点2667个。通过结果分析，预测槲皮素、木犀草素、山柰酚等有效成分通过IL6、TP53、IL1β等关键靶点，参与了细胞对脂质的反应等生物学过程，与Pathways in cance等通路密切相关。分子对接可知，槲皮素、木犀草素、山柰酚与IL6、TP53、IL1β具有较好的亲和力。体内实验证实马齿苋可显著抑制小鼠结肠肿瘤的发生，并抑制IL6、IL1β炎症因子水平及提高P53蛋白的表达水平。体外实验表明，槲皮素、木犀草素、山柰酚可不同程度抑制HCT116细胞增殖，木犀草素可抑制IL6、IL1β表达，上调P53表达。结论 通过网络药理学和体内外实验，证实了马齿苋治疗结直肠癌"多成分-多靶点-多通路"的作用特点，揭示其抑制炎症进而延缓炎癌转化可能是其潜在的治疗途径之一，为其进一步的机制探索和临床应用提供了理论依据和研究方向。
Network-Based Pharmacology and Experimental Validation to Explore the Portulaca oleracea L.Mechanism of Action in the Treatment of Colorectal Cancer
Objective To analyze the potential mechanism of action of Portulaca oleracea L.in the treatment of colorectal cancer(CRC)using network pharmacology and experimental verification.Methods With reference to TCMSP and ETCM databases,the effective ingredients and targets of Chinese medicine P.oleracea were obtained,and targets related to CRC were retrieved from Genecards,Drugbank and OMIM databases,which were imported into Venny database,String database and Cytoscape platform to construct the PPI proteins.The common core targets were analyzed for GO function and KEGG pathway enrichment using Metascape database and Microbiology Letter platform,and the active ingredients and key targets were molecularly docked using Autodock and Pymol software.To construct a CRC mouse model and give P.oleracea treatment,observe the histopathological changes of its colon,and validate the key targets using qRT-PCR experiments and Western blot experiments.In addition,quercetin,luteolin,and kaempferol were used to act on HCT116 cells to observe the effects on cell proliferation and calculate the IC50,which was further verified using qRT-PCR assay and Western blot assay.Results Network pharmacological yielded 11 active ingredients and 209 targets corresponding to the target of P.oleracea.Disease targets were 2667.By analyzing the results,it was predicted that the active ingredients such as quercetin,luteolin,and kaempferol were involved in biological processes such as cellular response to lipids through key targets such as IL6,TP53,and IL1β,which were closely related to pathways in cance.Molecular docking showed that quercetin,luteolin,and kaempferol had good affinity for IL6,TP53 and IL1β.In vivo experiments confirmed that P.oleracea significantly inhibited colon tumorigenesis in mice,and inhibited the levels of IL6,IL1β inflammatory factors and increased the expression level of P53 protein.In vitro experiments showed that quercetin,luteolin and kaempferol could inhibit the proliferation of HCT116 cells to different degrees,and luteolin could inhibit the expression of IL6 and IL1β and up-regulate the expression of P53.Conclusion Through network pharmacology,in vivo and in vitro experiments,the"multi-component-multi-target-multi-pathway"effect of P.oleracea in the treatment of CRC has been confirmed,which reveals that inhibition of inflammation and delay of inflammation-cancer transformation may be one of the potential therapeutic pathways,and provides a theoretical basis and research direction for further exploration of the mechanism and clinical application of P.oleracea.
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