基于网络药理学和动物实验探究羟基红花黄色素A治疗肺纤维化的作用机制

Mechanism of hydroxysafflower yellow A in the treatment of pulmonary fibrosis based on network pharmacology and animal experiment

基于网络药理学及动物实验探讨羟基红花黄色素A(hydroxysafflor yellow A,HSYA)治疗肺纤维化(pulmonary fibrosis,PF)的作用靶点及机制。利用网络药理学相关数据库SwissTargetPrediction、PharmMapper、GeneCards、OMIM、TTD筛选HSYA和PF的作用靶点,取两者交集得到共同靶点,构建蛋白相互作用(protein-protein interaction,PPI)网络。借助DAVID数据库进行GO及KEGG富集分析,以探究HSYA治疗PF可能的分子机制。采用博来霉素建立肺纤维化小鼠模型,通过HE、Masson染色对肺组织进行形态学观察,并检测肺组织中羟脯氨酸(hydroxyproline,HYP)含量;运用免疫组化和Western blot对小鼠肺组织中磷脂酰肌醇3-激酶/蛋白激酶B/哺乳动物雷帕霉素靶蛋白(phosphatidylinositol 3-kinase/protein kinase B/mammalian target of rapamycin,PI3K/AKT/mTOR)通路的相关蛋白进行检测。网络药理学方法筛选出药物疾病的共同靶点132个;GO富集分析得到719个条目,KEGG通路富集分析筛选出160条信号通路,主要涉及PI3K-AKT、Ras、MAPK等信号通路。IHC和Western blot结果显示,HSYA组肺组织中的PI3K、AKT、mTOR的蛋白表达较模型组显著减少(P<0.01)。网络药理学分析及动物实验结果提示,HSYA可能通过调控PI3K/AKT/mTOR信号通路来影响PF的发病进程,为HSYA的临床应用提供了科学依据。

This study aims to investigate the targets and mechanisms of hydroxysafflower yellow A(HSYA)in the treatment of pulmonary fibrosis(PF)based on network pharmacology and animal experiments.Network pharmacology databases such as SwissTarget Prediction,PharmMapper,GeneCards,OMIM,and TTD were used to screen the targets of HSYA and PF,common target and protein-protein interaction(PPI)network were obtained by the intersection.Using the DAVID database for GO and KEGG enrichment analysis,to explore the possible molecular mechanisms of HSYA treatment for PF.The mouse model of pulmonary fibrosis was established by bleomycin.The morphology of lung tissue was observed by HE and Masson staining,and the content of hydroxyproline(HYP)in lung tissue was detected;Immunohistochemistry and Western blot were used to detect the proteins related to phosphatidylinositol 3-kinase/protein kinase B/mammalian target of rapamycin(PI3K/AKT/mTOR)pathway in mouse lung tissue.The network pharmacology method screened 132 common targets for drug diseases;GO enrichment analysis yielded 719 entries while KEGG pathway enrichment analysis screened 160 signaling pathways,mainly involving PI3K-AKT,Ras,MAPK and other signaling pathways.IHC and Western blot results showed that the protein expression of PI3K,AKT,and mTOR in the lung tissue of the HSYA group was significantly reduced compared to the model group(P<0.01).Network pharmacology analysis and animal experimental results suggest that HSYA may affect the pathogenesis of PF by regulating the PI3K/AKT/mTOR signaling pathway,providing scientific basis for the clinical application of HSYA.