Label-free plasma proteomics reveals the regulatory mechanism of Chaige Qingsan Decoction on complement and interferon pathways against influenza.
CGQSD shows potential in modulating immune pathways in influenza, but clinical relevance requires further investigation.
Where it sits
this study against the rest of the vip (vasoactive intestinal polypeptide) corpusSummary and findings
This study explored the effects of Chaige Qingsan Decoction (CGQSD) on influenza A virus-induced respiratory infections using plasma proteomics and ELISA validation. In a cohort of 22 patients, 15 received CGQSD monotherapy, 4 received oseltamivir, and 3 received combination therapy. CGQSD modulated 178 differentially expressed proteins in complement and interferon pathways, with significant regulation of C4BPB and IFI6 confirmed by ELISA.
Abstract
This study investigates the material basis and mechanism of Chaige Qingsan Decoction (CGQSD) in early influenza A virus-induced respiratory infections using plasma proteomics and enzyme-linked immunosorbent assay (ELISA) validation. Ultra-performance liquid chromatography coupled with Orbitrap high-resolution mass spectrometry (UPLC-Orbitrap HRMS) characterized the chemical profile of CGQSD. In the discovery cohort, 54 plasma samples from 22 influenza A patients (paired pre- and post-treatment) and 5 healthy controls underwent label-free quantitative proteomics. Patients received CGQSD monotherapy (n = 15), oseltamivir alone (n = 4), or combined drug therapy (n = 3). Key candidate proteins were validated via ELISA in an independent cohort (76 samples from 32 patients and 6 controls). A total of 1444 proteins were quantified after removing those with > 75% missing values, with high technical reproducibility (quality control [QC] r > 0.98). Compared with healthy controls, 263 differentially expressed proteins (DEPs) were identified. CGQSD modulated 178 DEPs enriched in complement/coagulation and interferon pathways. ELISA confirmed the significant regulation of C4BPB (P = 0.0003) and IFI6 (P = 0.016), consistent with proteomics.CGQSD mediates anti-influenza activity through a dual immunomodulatory mechanism centered on "Complement-Interferon Homeostasis," targeting C4BPB (a complement regulator) and IFI6 (an interferon-stimulated gene). CGQSD monotherapy fine-tunes complement activation via precise C4BPB downregulation; combination therapy buffers this effect and normalizes the interferon pathway by universally decreasing IFI6. Notably, combination therapy triggers extracellular matrix remodeling (e.g., COL1A upregulation), indicating enhanced tissue repair beyond antiviral effects. These findings provide a molecular foundation for CGQSD and advocate for its further development as a host-directed immunomodulatory therapy.
Background
The study addresses the need to understand the molecular mechanisms by which Chaige Qingsan Decoction (CGQSD) exerts its effects on influenza A virus-induced respiratory infections. Previous research has suggested that CGQSD may have immunomodulatory properties, but the specific pathways and proteins involved were not well characterized. This study aims to elucidate these mechanisms using advanced proteomics techniques.
Methods
The study utilized a label-free quantitative proteomics approach on plasma samples from 22 influenza A patients and 5 healthy controls. Patients were treated with CGQSD monotherapy, oseltamivir, or combination therapy. Proteins were quantified using UPLC-Orbitrap HRMS, and key findings were validated with ELISA in an independent cohort of 32 patients and 6 controls.
Results
The primary finding was the modulation of 178 differentially expressed proteins by CGQSD, specifically in complement and interferon pathways. ELISA validation confirmed significant regulation of C4BPB (P=0.0003) and IFI6 (P=0.016). Combination therapy also indicated extracellular matrix remodeling, suggesting enhanced tissue repair.
Interpretation
The study provides evidence that CGQSD modulates immune pathways, potentially offering a host-directed therapy for influenza. However, the clinical significance of these findings remains uncertain due to the small sample size and reliance on proteomics data. Further research is needed to confirm these effects in larger, more diverse populations.
Key findings
- 1444 proteins quantified after filtering.
- 263 differentially expressed proteins identified vs controls.
- 178 DEPs modulated by CGQSD in complement/interferon pathways.
- C4BPB regulation confirmed by ELISA, P=0.0003.
- IFI6 regulation confirmed by ELISA, P=0.016.
Limitations
- small n=22 in primary cohort
- surrogate endpoints used
- proteomics data may not translate to clinical outcomes
- short-term follow-up
- single-site study