Network pharmacology and in vitro study of Fengreqing oral liquid(风热清口服液) in the intervention of wind-heat pattern

OBJECTIVE:To investigate the underlying mechanism of the effect of Fengreqing oral liquid(风热清口服液,FOL)on wind-heat pattern(WHP).METHODS:In this study,we predicted the potential targets of FOL via the approach of network pharmacology and verified it by in vitro inflammation model.In the network pharmacology part,two strategies,namely the direct target search and the indirect one,were used to collect the target sets of FOL in WHP treatment.The enrichment analysis was carried out by David database and Clue Go plug-in in Cytoscape.Furthermore,the potential targets were mapped in the candidate pathways.In the verification experiment section,in vitro model of lipopolysaccharide(LPS)induced RAW 264.7 was used to confirm the predictive results in the network pharmacology part.RESULTS:Through the two screening strategies,a total of 141 non-repetitive intervention targets of FOL on WHP were obtained.Gene ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)enrichment analysis showed that the intervention effect was mainly focused on the anti-inflammatory effect,and the Toll-like receptor signaling pathway was one of the most critical regulatory pathways.Further mapping analysis showed that phosphatidylinositol 3-kinase(PI3 K)-protein kinase B(AKT)signaling transfer might be the key part of regulating the concentration of inflammation mediators of FOL in the Toll-like receptor signaling pathway.In vitro experiment showed that FOL significantly reduced the levels of NO,IL-1,IL-6,and TNF-αproduced by RAW264.7 induced by LPS.Further immunofluorescence found that this effect is related to the regulation of PI3 K-AKT pathway activity by FOL.CONCLUSION:FOL can intervene in WHP by regulating the content of inflammatory mediators via the PI3 K-AKT pathway.