Ligustilide and tectoridin from Danggui Buxue Decoction synergistically attenuate atherosclerosis by targeting the CD36/TRPM2 axis and TNF-α/NF-κB pathway.

BACKGROUND: Atherosclerosis (AS) is a common chronic disease and the primary pathological basis for myocardial infarction, stroke, and other disabling conditions. Danggui Buxue Decoction (DBD) is a classic formula in Traditional Chinese Medicine (TCM) normally used for deficiency patterns. However, whether DBD can alleviate atherosclerosis, which bioactive constituents are responsible, and which molecular mechanisms are involved remain to be further investigated. PURPOSE: This study aims to characterize the absorbed bioactive components of DBD and decipher the multi-target mechanisms by which it counteracts atherosclerosis, providing a scientific rationale for its clinical application. METHODS: This study employs an integrated approach combining network pharmacology, WGCNA, and machine learning screening strategies, supported by experimental validation, to systematically elucidate the active pharmaceutical ingredients and molecular mechanisms of anti-AS effects of serum-absorbed constituents of DBD after oral administration. RESULTS: Network pharmacology and machine learning analyses predicted CD36, TRPM2, and TNF-α as three core targets for DBD intervention in AS. CETSA and BLI experiments further confirmed stable binding of Ligustilide and Tectoridin in DBD to these targets. In vivo and in vitro experiments demonstrated that the combined treatment of Ligustilide and Tectoridin effectively inhibit macrophage foaming, mitigate oxidative damage, and downregulate inflammatory cytokine release. CONCLUSION: This study successfully identified Ligustilide and Tectoridin as the primary bioactive constituents of DBD against atherosclerosis. By modulating CD36, TRPM2, and TNF-α, these compounds synergistically inhibit foam cell formation and inflammation, offering a novel pharmacological perspective on the anti-AS efficacy of DBD.