基于网络药理和细胞实验探索芪玉三龙方治疗非小细胞肺癌的机制

Mechanism of Action of Qiyu Sanlong Decoction in Treatment of Non-small Cell Lung Cancer Based on Network Pharmacology and Cell Experiments

目的采用网络药理学和细胞实验探究芪玉三龙方治疗非小细胞肺癌的潜在作用机制。方法利用中药系统药理学数据库与分析平台和中医药整合数据库检索芪玉三龙方成分筛选靶点,通过美国肿瘤基因组图谱数据库获得肺癌靶点基因。构建蛋白相互作用网络,并采用基因本体数据库和京都基因和基因组百科全书数据库进行富集分析。对核心靶蛋白与化合物进行分子对接分析,并以A549细胞进行实验验证。结果芪玉三龙方对于非小细胞肺癌的潜在有效靶点162个,蛋白相互作用网络分析得到核心靶点基因20个,主要的生物学过程为对缺氧的反应、MAPK级联、炎症反应,主要的功能通路为钙信号通路、TNF信号通路、IL-17信号通路、细胞周期。分子对接结果表明,MMP9-槲皮素、PPARG-异鼠李素、PTGS2-刺芒柄花素、JUN-刺芒柄花素之间存在有效的结合亲和力。在A549细胞实验中,PCR和Western blot分析结果表明,芪玉三龙方含药血清能够促进PPAR-γmRNA和蛋白表达(P<0.05),抑制c-JUN、COX-2、MMP-9 mRNA和蛋白表达(P<0.05)。结论芪玉三龙方通过影响A549肺癌细胞的凋亡、侵袭和肿瘤血管生成及炎症进程达到干预肺癌的作用。

Objective To investigate the potential mechanism of action of Qiyu Sanlong Decoction in the treatment of non-small cell lung cancer based on network pharmacology and cell experiments.Methods TCMSP and TCMID databases were searched to obtain the chemical components and targets of Qiyu Sanlong Decoction,and TCGA database was analyzed to obtain the gene targets of lung cancer.A protein-protein interaction(PPI)network was constructed,and gene ontology and Kyoto Encyclopedia of Genes and Genomes were used for enrichment analysis.Molecular docking was performed for core target proteins and compounds,and A549 cells were used for experimental validation.Results Qiyu Sanlong Decoction had 162 potential effective targets in the treatment of non-small cell lung cancer.The PPI network analysis obtained 20 core target genes.The main biological processes included response to hypoxia,MAPK cascade,and inflammatory response,and the main functional pathways included the calcium signaling pathway,the TNF signaling pathway,the IL-17 signaling pathway,and cell cycle.Molecular docking showed effective binding affinity between MMP9 and quercetin,between PPARG and isorhamnetin,between PTGS2 and formononetin,and between JUN and formononetin.In A549 cell experiments,PCR and Western blot showed that serum containing Qiyu Sanlong Decoction could promote the mRNA and protein expression of PPAR-γ(P<0.05)and inhibit the mRNA and protein expression of c-JUN,COX-2,and MMP-9(P<0.05).Conclusion Qiyu Sanlong Decoction exerts an interventional effect on lung cancer by affecting the apoptosis,invasion,angiogenesis,and inflammation of A549 cells.