Tongxinluo capsule attenuates atherosclerosis-related inflammation, oxidative stress, and endothelial dysfunction: Integrated component profiling, network pharmacology, and experimental validation

Introduction Tongxinluo (TXL) capsule, composed of 12 Chinese herbal extracts, is indicated for the treatment of coronary heart disease. This study aimed to elucidate the chemical composition of TXL capsules and uncover their potential mechanisms against atherosclerosis through integrated component profiling, network pharmacology, and experimental validation. Methods TXL constituents were systematically characterized using ultra-performance liquid chromatography-mass spectrometry (UPLC-MS). Network pharmacology was employed to predict therapeutic targets, followed by molecular docking. The anti-inflammatory activities of TXL were evaluated by measuring the inhibition of nitric oxide (NO), interleukin (IL)-6, and tumor necrosis factor (TNF)-α production in lipopolysaccharide (LPS)-induced RAW 264.7 cells. The ability of TXL to reduce oxidative stress was assessed by the nuclear factor erythroid 2-related factor 2 (Nrf2) luciferase and intracellular reactive oxygen species (ROS) assays. Finally, the capacity of TXL extract to restore endothelial cell survival and function was evaluated in human endothelial EA.hy926 cells using cell migration and tube formation assays. Key signaling pathways were validated through Western blotting and immunofluorescence as guided by network pharmacology and molecular docking. Results Based on the 97 active ingredients identified in TXL, network pharmacology revealed 340 compound-disease intersection targets enriched in inflammation, oxidative stress, endothelial dysfunction, and lipid-related pathways with strong binding affinities between major TXL components and core atherosclerosis-related targets. TXL dose-dependently reduced LPS-induced NO, IL-6, TNF-α, and ROS levels. In endothelial cells, TXL attenuated H 2 O 2 -induced oxidative injury, restored viability, decreased ROS accumulation, and significantly enhanced migration and tube formation. The molecular mechanisms of TXL were associated with suppression of NF-κB p65 nuclear translocation and activation of the Nrf2-HO-1 axis. Discussion TXL restored endothelial survival and function, which were at least partly attributed to modulation of the NF-κB inflammatory pathway and activation of the Nrf2-HO-1 antioxidant axis to suppress inflammation and oxidative stress. These findings underscore TXL’s potential as a promising adjunctive therapy for the treatment of atherosclerosis.