基于整合药理学模式探究小青龙汤防治高原肺水肿的作用机制OA

Objective:To explore the potential mechanism of Xiaoqinglongtang(XQL)in the prevention and treatment of high altitude pulmonary edema(HAPE)by network pharmacology,molecular docking,and molecular dynamics simulation,and to verify it by in vivo animal model.Methods:In this study,the active ingredients,drug targets,and HAPE-related targets of XQL were collected from BATMAN-TCM,GeneCards,and Online Mendelian Inheritance in Man(OMIM)databases.The protein-protein interaction(PPI)network was constructed by using intersection targets,and the core targets were screened and visualized by Cytoscape software.Functional annotation and pathway analysis of the intersection targets were performed by gene ontology(GO)and Kyoto encyclopedia of genes and genomes(KEGG)functional enrichment.AutoDock and GROMACS were used to evaluate the binding ability of active ingredients to key targets.In the experimental verification part,a mouse model of HAPE induced by hypobaric hypoxia(simulated 6 000 m altitude for 48 h)was established.The control effect was evaluated by hematoxylin-eosin(HE)staining,lung tissue water content,lung tissue wet/dry weight ratio,real-time quantitative polymerase chain reaction(Real-time PCR)detection of gene expression levels,and immunohistochemistry and Western blot detection of key protein expression.Results:A total of 355 active ingredients of XQL,2 142 targets,716 HAPE-related targets,and 236 intersection targets were obtained by network pharmacology analysis.Key core targets such as interleukin(IL)-6,tumor necrosis factor(TNF),protein kinase B1(Akt1),and hypoxia-inducible factor-1α(HIF-1α)were screened.The results of GO analysis of common targets involved 738 biological processes(BP),72 cellular components(CC),and 135 molecular functions(MF).KEGG analysis effectively enriched two important signaling pathways:Phosphoinositol 3-kinase(PI3K)/Akt and HIF-1α.The results of molecular docking and molecular dynamics simulation showed that the screened active ingredients had good binding ability with key targets.In the HAPE model induced by hypobaric hypoxia(6 000 m,48 h),the lung tissue water content,lung tissue wet/dry weight ratio,and pathological injury score of the model group were significantly increased(P＜0.01),accompanied by exudation of a large number of red blood cells in the alveoli and alveolar interstitium,a significant increase in inflammatory cells,a significant widening of the alveolar septum,and mutual fusion between the alveoli.The XQL administration group significantly improved the above pathological changes(P＜0.01).The results of inflammatory factor expression showed that compared with the control group,the model group showed significantly up-regulated expression of TNF-α,IL-6,and IL-1β in the lung tissue(P＜0.01).Compared with the model group,the XQL administration group had significantly decreased expression of inflammatory factors(P＜0.05,P＜0.01).The mRNA expression of key pathway related genes PI3K,Akt1,mammalian target of rapamycin(mTOR),and HIF-1αwas significantly increased in the model group(P＜0.01),and decreased in a concentration-dependent manner after XQL administration(P＜0.05,P＜0.01).The expression levels of key proteins PI3K,phosphorylation(p)-PI3K,Akt1,p-Akt1,mTOR,p-mTOR,and HIF-1α in lung tissue were analyzed by immunohistochemistry and Western blot.Compared with the blank group,the model group showed increased expression of key proteins(P＜0.05,P＜0.01).Compared with the model group,the XQL administration group exhibited decreased expression of key proteins(P＜0.05,P＜0.01).Conclusion:XQL can reduce lung inflammation and improve HAPE.The mechanism may be related to the regulation of PI3K/Akt/mTOR and HIF-1α pathways.This study provides a new idea and a theoretical basis for the treatment of HAPE with XQL.