Based On The IL-17 Signaling Pathway To Explore The Mechanism Of Chaishi Tuire Particle Infection With Influenza A Virus Was Explored

Background: Developed from a Traditional Chinese Medicine(TCM)classic famous prescription,Chaishi-Tuire Particle(CSTRP)has been widely used for treating influenza virus infection and showed the obvious therapeutic effect.However,little is known about the mechanism of CSTRP against influenza virus infection.Purpose: The goal of this study is to demonstrate the anti-inflammatory activity of CSTRP against the influenza A virus and its potential effect in regulating host immune response.Methods: A series of bioinformatics analyses including network pharmacology and molecular docking were firstly carried out to predict the main active compounds of CSTRP and the potential anti-inflammatory targets and related-signaling pathways.The antiinflammatory activity of CSTRP was determined by using the lipopolysaccharide(LPS)-induced macrophages RAW264.7 cells in vitro,the expressions of TNF-α and IL-6downstream inflammatory factors in IL-17 pathway were detected by ELISA and the underlying mechanism of the expression of important proteins TRAF6,and MAPK14 related m RNA and proteins were further analyzed by real-time quantitative PCR and Western Blotting.Influenza-associated mice pneumonia model was used to explore the anti-inflammatory and immunomodulatory activity of CSTRP in vivo,in which sick signs,body weight change,viral titers,the pathological parameters,and the secretion of inflammatory cytokines after CSTRP treatment was systematically determined.Results: Bioinformatics analysis predicted that six active compounds of CSTRP including quercetin,kaempferol,luteolin,beta-sitosterol,sitosterol,and stigmasterol could contribute to play a crucial part in the IL-17 signaling pathway.The following research confirmed that CSTRP significantly inhibited pro-inflammatory cytokines(TNF-α and IL-6)by suppressing the expression of TRAF6 and MAPK14 in LPS-stimulated macrophages RAW264.7 cells.Moreover,CSTRP treatment markedly improves the sick signs,weight loss,lung index,and lung pathological changes.Additionally,CSTRP administration decreased the virus load in lung tissues and reduced the secretion of TNF-α and IL-6 in lung tissues and serum of infected mice.Conclusion: CSTRP not only showed anti-inflammatory activity in vitro by down regulating the IL-17 signaling pathway but also exhibited a significant protective effect against lethal influenza A infection in vivo.Further studies showed that the in vivo antiinflammatory effect of CSTRP may be attributed to suppressing the expression of inflammatory cytokines and restricting viral replication.These findings provide evidence for clinical treatment of influenza A virus infection with CSTRP.