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

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.