Modulation of NF-κB and cytokine signaling by Cordia dichotoma and Cordia myxa attenuates methotrexate-induced pulmonary inflammation and oxidative injury in rats

Background Methotrexate (MTX) is a widely used anticancer and immunosuppressive agent, but its clinical use is limited by lung injury associated with oxidative stress, inflammation, and fibrosis. Leaves of Cordia dichotoma and Cordia myxa traditionally have been used for respiratory disorders, suggesting potential anti-inflammatory benefits. Objective To characterize the chemical profile of defatted aqueous methanol extracts (DAME) of C. dichotoma and C. myxa leaves and evaluate their protective effects against MTX-induced pulmonary inflammation and injury. Methods Extracts were analyzed by HPLC/MS. Rats were divided into control, MTX, Cordia -pretreated (250 or 500 mg/kg), and dexamethasone groups. Lung tissues were assessed for oxidative stress (MDA, GSH, SOD), inflammatory mediators (TNF-α, IL-1β, IL-6), NF-κB activation, fibrotic markers (TGF-β1, α-SMA, hydroxyproline), and histopathology. In vitro , RAW 264.7 macrophages were used to assess cytotoxicity, nitric oxide production, and NF-κB modulation. Results HPLC/MS identified 81 and 29 metabolites in C. dichotoma and C. myxa , respectively, mainly phenolic acids and flavonoids. MTX induced oxidative and inflammatory damage, with elevated cytokine levels and NF-κB activation. Pretreatment with both extracts, particularly C. dichotoma (500 mg/kg), significantly restored antioxidant balance, suppressed TGF-β1/α-SMA signaling, and preserved lung architecture. In vitro , the extracts inhibited LPS-induced NO release and NF-κB activation, indicating direct macrophage-modulating activity. Conclusion Phenolic-rich extracts of C. dichotoma and C. myxa exert anti-inflammatory, antioxidant, and anti-fibrotic effects by modulating macrophage-driven inflammation and restoring the pulmonary microenvironment, supporting their potential as a natural remedy for inflammation-associated lung injury.