基于网络药理学和实验验证探究麦味养肺汤通过调控线粒体自噬抗肺纤维化的作用机制

Mechanism of Maiwei Yangfei Decoction in the Treatment of Pulmonary Fibrosis through Regulation of Mitophagy Based on Network Pharmacology and Experimental Verification

目的:基于网络药理学和实验验证探究麦味养肺汤治疗肺纤维化的潜在作用靶点和机制。方法:利用网络药理学的方法获取疾病和药物的公共靶点,制作网络图和蛋白质相互作用(protein-protein interaction,PPI)网络图,并进行拓扑学分析;同时对核心靶点在基因本体(Gene Ontology,GO)和京都基因与基因组百科全书数据库(Kyoto Encyclopedia of Genes Genomes,KEGG)进行富集分析;最后通过病理切片染色、Western Blot、q-PCR试验验证网络药理学富集分析结果。结果:共筛选出19个关键靶点,包括OPTN、TNF、IL-6、MAPK8、PINK1等,富集分析结果显示差异性最大的是线粒体自噬信号通路以及细胞衰老信号通路,PTEN诱导的假定激酶1(PTEN-induced putative kinase 1,PINK1)、Parkin和P62是其潜在作用靶点,试验结果显示麦味养肺汤能明显改善肺组织炎症评分、纤维化评分(P<0.05或P<0.01),明显调控PINK1、P62、P53、MDM2蛋白表达,调控Pink1、P62、Tp53、Park2、Casp3 mRNA表达(P<0.05或P<0.01)。结论:麦味养肺汤可能是通过调控线粒体自噬和细胞衰老等多途径多靶点的方式发挥抗肺纤维化作用。

Objective:To explore the potential functional targets and mechanism of Maiwei Yangfei Decoction in the treatment of pulmonary fibrosis based on network pharmacology and experimental verification.Methods:Network pharmacology was used to obtain the common targets of the disease and the drug.Network diagram and protein-protein interaction(PPI)were plotted for topological analysis.The core targets were analyzed by Gene Ontology(GO)and Kyoto Encyclopedia of Genes Genomes(KEGG)enrichment.The results of network pharmacology enrichment analysis were verified by pathological section staining,Western blot,and q-PCR.Results:Nineteen key targets were screened out,including OPTN,TNF,IL-6,MAPK8,and PINK1.Enrichment results showed that the most significant difference was observed in the mitochondrial autophagy signaling pathway and cell senescence signaling pathway.PINK1,parkin,and P62 were potential targets.The experimental results showed that Maiwei Yangfei Decoction improved inflammatory score and fibrosis score of lung tissues(P<0.05 or P<0.01),regulated the protein expression of PINK1,P62,P53,and MDM2,and mRNA expression of Pink1,P62,Tp53,Park2,and Casp3(P<0.05 or P<0.01).Conclusion:Maiwei Yangfei Decoction may play an anti-pulmonary fibrosis role through regulating mitophagy and cell senescence by multi-channel and multi-target way.