[Integrating UPLC-Q-TOF-MS/MS and network pharmacology to explore serum and synovium migration components of Huangqin Qingre Chubi Capsules and potential mechanism of treating rheumatoid arthritis].

Huangqin Qingre Chubi Capsules(HQC) are commonly used in clinical practice to treat rheumatoid arthritis(RA). It is composed of Scutellariae Radix, Gardeniae Fructus, Coicis Semen, Clematidis Radix et Rhizoma, and the stir-fried Persicae Semen. However, the pharmacological substance basis and mechanism are not yet clear. This study systematically elucidated the pharmacological substance basis and mechanism of HQC by the research strategy of "identification of target tissue migration components-network mechanism prediction-multidimensional experimental verification". An adjuvant-induced arthritis(AA) rat model was established, and the serum and synovium migration components of HQC in normal and AA model rats were analyzed using ultra-high performance liquid chromatography-quadrupole time-of-flight mass spectrometry(UPLC-Q-TOF-MS/MS). These serum and synovium migration components were integrated for network pharmacology prediction, which was followed by multi-dimensional validation using molecular docking, RT-qPCR, and pharmacodynamic evaluation. The results showed that 46 serum and 13 synovium migration components were identified in normal rats, and 59 serum and 15 synovium migration components were identified in AA rats, indicating that the RA disease environment affects the entry of HQC into the bloodstream and migration to the synovium. Comparative analysis of components common and unique to normal and model groups identified geniposide, baicalin, daucosterol, coptisine, acteoside, luteolin, and crocin as migration components reaching the target site, reflecting the target tissue enrichment characteristics of HQC. Network pharmacology of the migration components screened 7 core targets: GAPDH, TNF-α, AKT1, PTGS2, NF-κB1, MAPK1, and SphK1, which were enriched in key RA signaling pathways including PI3K-AKT, HIF-1, VEGF, and TNF. Molecular docking and RT-qPCR confirmed good binding activity and regulatory capacity between key components and core targets. In vivo validation further demonstrated that HQC significantly improved joint inflammation and synovial hyperplasia in AA rats. This study confirmed that HQC exerted an anti-RA effect by targeting chronic inflammation and metabolic disorders through multi-target regulation of PI3K-Akt and other signaling pathways via serum and synovium migration components. The research provides a scientific basis for the clinical application of HQC and offers a new strategy for studying the mechanisms of TCM.