Study On The Antipyretic And Anti-inflammatory Effects And Mechanism Of Huanglian Jiedu Decoction On Lipopolysaccharide Induced Fever In Rats

Aim and Significance:In this study,Huanglian Jiedu Decoction(HLJDD),a traditional Chinese medicine prescription,was taken as the research object,guided by traditional Chinese medicine theories,pharmacology and molecular biology techniques as research methods,combined with network pharmacology,molecular docking,metabolomics and other research methods,to comprehensively investigate the antipyretic effect and potential mechanism of HLJDD on rat fever model induced by LPS.The aim is to provide reference for the antipyretic effect research and application development of HLJDD.Methods:①TCMSP,BATMAN-TCM and ETCM platforms were used to search the main components and protein targets of HLJDD,and Cytoscape3.8.2 was used to construct the component and target network.Disease targets were obtained from GeneCard,DisGeNET and DrugBank platforms,and the overlapping targets of ingredients and diseases were imported into STRING database to construct PPI network.The core target was analyzed by GO and KEGG,and then molecular docking was carried out between the core target and the component.②LPS was used to replicate rat fever model and the changes of anal temperature were monitored.The expressions of TNF-α,IL-6 and IL-1β in serum,and the expressions of PGE2 and cAMP in hypothalamus were determined by ELISA.The expression of MAPK-related proteins predicted by network pharmacology was detected by Western Blotting.③Metabolomics technology was used to detect the changes of serum metabolites in rats,screen potential differential metabolites,and enrich related signaling pathways to elucidate the metabolic characteristics of antipyretic effect of HLJDD.Results:①The results of network pharmacology showed that TNF,JUN,AKT1,TP53,IL6 and IL-1β were the potential key proteins in the antipyretic and antiinflammatory effects of HLJDD.The results of enrichment analysis suggested that the signaling pathways involved in the regulation of antipyretic and anti-inflammatory effects of HLJDD involved MAPK signaling pathway,TNF signaling pathway and Toll-like receptor signaling pathway.The major ingredients of HLJDD could stably bind to TNF,JUN,and AKT1 proteins,according to the results of molecular docking.② Pharmacological effect investigation showed that the body temperature of rats increased significantly after LPS modeling,and HLJDD could improve the disorder of body temperature regulation,regulate the expression levels of TNF-α,IL-6,IL-1β,PGE2 and cAMP biochemical indexes.The results of molecular biology study showed that the expression levels of ERK,JNK and p3 8 phosphorylated proteins in hypothalamus tissues of rats were reduced in different dose groups of HLJDD.③The results of metabolomic study showed that seven differential endogenous metabolites were initially identified from rat serum under positive and negative ionization mode of electrospray ion source.The enrichment analysis of signaling pathways showed that Glycerophospholipid metabolism,Arachidonic acid metabolism,Phenylalanine,tyrosine and tryptophan biosynthesis,Phenylalanine metabolism were highly correlated with the metabolic regulation of HLJDD.Conclusions:The results of this study showed that HLJDD can significantly improve the fever symptoms induced by LPS,regulate the expression of biochemical indexes,and its mechanism is related to the inhibition of MAPK signaling pathway.Combined with metabolomics,it was found that HLJDD could affect different metabolic pathways and slow down the occurrence of inflammation.This study preliminarily explained the antipyretic and anti-inflammatory pharmacological action and molecular mechanism of HLJDD,which provided experimental basis and theoretical support for further research.