Immunomodulatory mechanism of Huangqi-Guizhi-Wuwu Decoction in alleviating autoimmune arthritis based on network pharmacology and experimental validation.

BACKGROUND

Juvenile idiopathic arthritis (JIA) is the most common type of childhood autoimmune arthritis. Huangqi Guizhi Wuwu decoction (HGWD), a traditional Chinese herbal formula, is widely used in China to treat patients with autoimmune arthritis. However, the bioactive ingredients and their complex regulatory mechanisms remain unclear.

PURPOSE: To investigate the active components of HGWD using a novel comprehensive strategy and clarify the mechanism underlying immunomodulation.

METHODS

The main active components of HGWD were determined using ultra-high-performance liquid chromatography-high resolution mass spectrometry (UPLCHRMS). The core target and biological immune regulation mechanism of HGWD in alleviating JIA were predicted using combined network pharmacology and molecular docking analyses, followed by in vitro and in vivo experiments.

RESULTS

A total of 1387 active components were identified by UPLC-MS, of which eight were the main active ingredients. Network pharmacology showed that HGWD acted on core targets, such as STAT3. Further combined analysis revealed that regulation of the Th17 differentiation pathway may be an important mechanism by which HGWD relieves JIA. Molecular docking verification showed that the key component of HGWD can stably bind JAK/STAT-related proteins. The induced differentiation of Th17 and Treg in vitro experiment confirmed the immunoregulatory effects of HGWD. in vivo experiments, HGWD significantly alleviated symptoms of arthritis in a mouse model of collagen-induced arthritis (CIA) and was closely associated with restoring the Th17/Treg balance.

CONCLUSION

Taken together, serum components/UPLC-MS, network pharmacology, and molecular biology analyses are feasible strategies for exploring the active ingredients in HGWD. This study highlights the clinical potential of HGWD in alleviating JIA and provides evidence of its therapeutic potential through immune regulation.