镰形棘豆总黄酮抗肾癌分子作用机制的网络药理学初探

Preliminary study on the molecular mechanism of total flavonoids of Oxytropis falcata anti renal cell carcinoma by network pharmacology

目的利用网络药理学和分子对接方法研究镰形棘豆总黄酮(TFOF)抗肾癌的分子机制。方法通过SEAdatabase、SwissTarget Prediction和TargetNet数据库预测TFOF的相关靶点。从DisGeNET、NCBI、GeneCards、OMIM和GeneCLip3数据库收集与肾癌相关的靶点,通过靶点映射确定活性化合物作用于炎症的靶点。利用Cytoscape3.7.2软件,构建药物-化合物-疾病-靶点蛋白互作网络图。借助Omicshare和Metascape开放平台对交集靶点进行GO及KEGG富集分析。检测活性化合物对TNF-α诱导特殊转染的人胚胎肾293细胞(HEK-293细胞)中NF-κB抑制率的影响,以验证其抗肿瘤活性。通过Autodock软件对核心化合物和核心靶点进行分子对接验证。结果靶点映射确定了460个镰形棘豆黄酮类化合物与炎症的共同靶点,其中,关键靶点8个(包括AKT1、TNF、SRC、VEGFA、EGFR等)。KEGG富集结果显示:TFOF的抗炎机制主要为调控cAMP信号通路、癌症中的微小RNA、VEGF信号通路等。分子对接显示核心成分与核心靶点对接良好。体外实验表明:7-羟基黄酮、2′,4′-二羟基查尔酮这两种化合物可显著抑制TNF-α活化HEK-293细胞中NF-κB的释放,具有明显的抗肾癌作用。结论镰形棘黄酮类化合物通过“多成分-多靶点-多通路”发挥抗肾癌作用,该作用可能与其对cAMP信号通路、癌症中的微小RNA、VEGF信号通路等的调控有关。

OBJECTIVE To study the molecular mechanism of tolal flavonoids of Oxytropis falcata(TFOF)against renal cell carcinoma using network pharmacology and molecular docking methods.METHODS TFOFB-related targets was predicted through SEA database,SwissTarget Prediction and TargetNet database.The targets related to kidney cancer was collected from the databases of DisGeNET,NCBI,GeneCards,OMIM and GeneCLip3,and the targets of active compounds on inflammation were determined through target mapping.Cytoscape 3.7.2 software was used to construct a drug-compound-disease-target protein interaction network diagram.With the help of Omicshare and Metascape open platform,GO and KEGG enrichment analysis of the intersection target are carried out.We detected the effect of active compound on NF-κB inhibition rate in human embryonic kidney 293 cells(HEK-293 cells)induced by TNF-αto verify its anti-cancer activity.Autodock software was used to verify the molecular docking between the core compound and the core target.RESULTS Target mapping identified 460 common targets of falciparum flavonoids and inflammation,including 8 key targets,such as AKT1,TNF,SRC,VEGFA,EGFR,etc.KEGG enrichment results showed that TFOFB anti-inflammatory mainly regulates cAMP signaling pathway,microRNA in cancer,VEGF signaling pathway,etc.Molecular docking showed that the core components were well docked with the core targets.In vitro experiments showed that 7-hydroxyflavanone and 2′,4′-dihydroxychalcone could significantly inhibit the release of NF-κB in HEK-293 cells activated by TNF-α,and had obvious anti-renal cancer effects.CONCLUSION Sickle flavonoids play an anti-kidney cancer effect through“multi-component-multi-target-multi-pathway”,whose effect may be related to its regulation of cAMP signaling pathway,microRNA in cancer,VEGF signaling pathway,etc.