基于网络药理学探讨补阳还五汤治疗特发性肺纤维化的生物学机制

To explore the biological mechanism of Buyang Huanwu Decoction on idiopathic pulmonary fibrosis based on network pharmacology

目的 运用网络药理学研究方法预测补阳还五汤(BYHWT)治疗特发性肺纤维化(IPF)的潜在靶点及可能机制,并通过转化生长因子(TGF)-β1诱导肺泡上皮细胞A549的上皮间质转化(EMT)进行体外实验验证。方法 根据网络药理学的原理获得BYHWT治疗IPF的潜在作用靶点,利用Cytoscape软件绘图、STRING网络平台、Metascape数据库进行蛋白互作(PPI)网络、基因本体(GO)和京都基因与基因组百科全书(KEGG)富集分析,以TGF-β1诱导肺泡上皮细胞为体外模型,对网络药理学预测的信号通路进行实验验证。结果 网络药理学预测BYHWT治疗IPF的潜在作用靶点为168个,KEGG富集分析相关信号通路有磷脂酰肌醇-3-激酶/蛋白激酶B(PI3K/Akt)信号通路、核转录因子(NF)-κB信号通路及FOXO信号通路等。细胞实验结果证明,较模型组相比,BYHWT可显著下调p-Akt1/2/3、磷酸化NF-κB抑制蛋白(p-IκBα)的蛋白水平,并在RNA水平显著下调EMT相关标记蛋白Ⅰ型胶原(Coll)A1、锌指蛋白(Snail)和波形蛋白(Vimentin)的基因表达(P<0.05)。结论 BYHWT具有多成分、多靶点及多通路的特点防治IPF,其主要通过抑制PI3K/Akt/NF-κB的信号通路发挥治疗IPF的作用。

Objective To predict the potential targets and possible mechanisms of the Buyang Huanwu decoction(BYHWT)for the treatment of idiopathic pulmonary fibrosis(IPF)using a network pharmacology study,and to validate in vitro experiments by transforming growth factor(TGF)-β1-induced epithelial-mesenchymal transition(EMT)in alveolar epithelial cells A549.Methods According to the principle of network pharmacology,the potential targets of BYHWT in the treatment of IPF were obtained.Cytoscape software drawing,STRING network platform and metascape database were used for protein interaction(PPI)network,gene ontology(GO)and kyoto encyclopedia of genes and genomes(KEGG)enrichment analysis.TGF-β1-induced alveolar epithelial cells were used as the in vitro model.Experimental validation of network pharmacological prediction signaling pathway was carried out.Results The network pharmacology predicted 168 potential targets of BYHWT for the treatment of IPF,and the signaling pathways associated with KEGG enrichment analysis were PI3K/AKT signaling pathway,nuclear transcription factor(NF)-κB signaling pathway,and FOXO signaling pathway.Cell experiment results showed that compared with the model group,BYHWT could significantly down-regulate the protein levels of p-Akt1/2/3 and phosphorylated NF-κB inhibitor protein(p-IκBα),and the gene expressions of EMT-related marker proteins typeⅠcollagen(Coll)A1,snail and vimentin were significantly down-regulated at RNA level(P<0.05).Conclusions BYHWT has the characteristics of multi-component,multi-target and multi-pathway to preventing and curing IPF,which plays a main role in the treatment of IPF by inhibiting the PI3K/Akt/NF-κB signaling pathway.