Quantitative Proteomics Analysis of Systemic Responses and Biological Mechanisms of ShuFengJieDu Capsule Using H1N1-Infected RAW264.7 Cells

[Image: see text] Emerging infectious diseases (EIDs) are a significant burden on global economies and public health to any country in the world. With the extensive application of traditional Chinese medicines (TCMs) for EID treatment, the underlying molecular mechanisms have caught more attention than before. The ShuFengJieDu capsule (abbreviated as SFJD) is a TCM prescription used for treating upper respiratory infection (URI) with symptoms of fever, sore throat, headaches, nasal congestion, and cough for more than 30 years in China. SFJD is also widely used for the prevention and treatment of viral infectious diseases, especially for the EIDs. In this study, a bioactivity-integrated method of ultraperformance liquid chromatography quadrapole/time-of-flight mass spectrometry combined with methyl thiazolyl tetrazolium assay was applied to screen potential antivirus compounds in SFJD on the H1N1-infected RAW264.7 cell models. Three compounds (forsythoside E, verbenalin, and emodin) exert the advantages of protective effects in cell vitality during H1N1 infection. The isobaric tags for relative and absolute quantification (iTRAQ)-coupled liquid chromatography coupled to tandem mass spectrometry (LC–MS/MS) analysis and the subsequent quantitative proteome analysis were performed to investigate the potential molecular mechanisms triggered by these three bioactive compound-triggered molecular mechanisms in H1N1-infected RAW264.7 cells. Dysregulated proteins were involved in regulating the levels of proinflammatory cytokines, the IFN (interferon)-stimulated gene signal in the Type I IFN, TBK/IRF3, and MAPK/NF-κB signaling pathways. In conclusion, we identified the main bioactive compounds in SFJD exerting antiviral effects and illuminated that Type I IFN and MAPK/NF-κB signaling pathways are involved in the anti-H1N1 infection effects of SFJD. Our study not only provides solid theoretical support for the clinic application of SFJD but also sheds light on the novel research methods for TCM study.