A defined three-compound combination from Artemisia halodendron inhibits PLA2 and 5-LOX and attenuates inflammatory...

Artemisia halodendron is a traditional Chinese medicine (TCM) with anti-inflammatory activity in clinical practice, yet its active constituents and underlying synergistic mechanism remain unclear. This study aimed to identify its anti-inflammatory components and develop a synergistic multi-compound formulation in a collagen-induced arthritis (CIA) rat model. Network pharmacology identified the leukotriene biosynthesis pathway as a critical axis, highlighting phospholipase A2 (PLA2) and 5-lipoxygenase (5-LOX) as therapeutic targets. Based on these predictions and enzymatic validation, a synergistic three-compound combination (SMG) consisting of isorhamnetin (G8), 6,7-dimethoxy-3',4',5-trihydroxyflavone (G1), and 7-O-methyleriodictyol (G16) was screened. Optimized SMG (molar ratio 2.7:1.0:8.8) inhibited PLA2 and 5-LOX at low micromolar levels. Functionally, in LPS-stimulated macrophages, SMG reduced Pla2g2a and Alox5 expression and significantly decreased LTB4 production, providing evidence for suppression of a representative 5-LOX-derived inflammatory lipid mediator. In 16HBE cells, SMG also attenuated TGF-β1-induced epithelial-mesenchymal transition (EMT) by downregulating profibrotic and proangiogenic programs. In vivo, oral SMG administration significantly reduced joint swelling and improved pathology in CIA rats, with efficacy comparable to the crude extract. This study thus defines SMG as a precise multi-target inhibitor, acting via dual PLA2/5-LOX blockade. These results provide a promising lead for chronic inflammatory disorders and a validated approach to advancing traditional herbal medicines.