Qingjin Pingchuan formula attenuates pulmonary inflammation by reprogramming neutrophil SHP1-JAK2/SRC-STAT3 signaling

The Qingjin Pingchuan Formula (QJPC), a multi-herb traditional Chinese medicine prescription, is clinically employed for severe community-acquired pneumonia (SCAP), though its underlying molecular mechanisms remain incompletely understood.

This study aimed to elucidate the anti-inflammatory effects of QJPC and its impact on neutrophil-mediated signaling pathways.

An integrative strategy combining network pharmacology, single-cell RNA sequencing (scRNA-seq), and experimental validation was applied. Chemical profiling of QJPC and drug-containing human serum was conducted using UPLC-MS. Network pharmacology and molecular docking analyses predicted and verified potential targets. A murine lipopolysaccharide (LPS)-induced acute lung injury model was established for in vivo studies. Functional assays were performed on primary neutrophils, and neutrophil-depleted mice were used to confirm neutrophil-dependent effects. The SHP1 agonist SC43 was co-administered to further assess SHP1-mediated mechanisms.

QJPC alleviated pulmonary inflammation by modulating the SHP1-JAK2/SRC-STAT3 axis in neutrophils. UPLC-MS identified 17 systemically absorbed constituents. Network pharmacology predicted JAK2, SRC, and STAT3 as key targets, which were supported by molecular docking. In LPS-injured mice, QJPC improved histopathology, reduced IL-6, and elevated IL-10 levels. scRNA-seq demonstrated that QJPC reshaped neutrophil heterogeneity and upregulated Ptpn6, encoding the phosphatase SHP1. Functional assays confirmed that QJPC enhanced SHP1 protein expression, suppressed STAT3 phosphorylation and nuclear translocation, and attenuated JAK2 and SRC activation in primary neutrophils. In vivo, QJPC's anti-inflammatory effects were diminished in neutrophil-depleted mice and modulated by SC43 co-treatment. Together, these findings indicate that QJPC exerts lung-protective effects, at least partly, through SHP1 upregulation and subsequent inhibition of the JAK2/SRC-STAT3 pathway in neutrophils. This study provides mechanistic insight into the anti-inflammatory action of QJPC and supports its therapeutic potential in neutrophil-driven pulmonary inflammatory diseases.