Zhongguo Zhong Yao Za Zhi. 2026 Jul;51(13):3761-3770. doi: 10.19540/j.cnki.cjcmm.20260305.705.
ABSTRACT
This study integrated network pharmacology, computational simulation, spatial transcriptomics, and animal experiments to systematically reveal the multi-level mechanism of the traditional Chinese medicine compound Zuopi Pills in treating atherosclerosis(AS) by inhibiting ferroptosis via the acyl-CoA synthetase long-chain family member 4(ACSL4)/lysophosphatidylcholine acyltransferase 3(LPCAT3)/arachidonate 15-lipoxygenase(ALOX15) signaling axis. Network pharmacology was used to screen for the intersecting targets between the active components of Zuopi Pills and AS, followed by protein-protein interaction(PPI) construction, Gene Ontology(GO) and Kyoto Encyclopedia of Genes and Genomes(KEGG) pathway enrichment analyses. Molecular docking and molecular dynamics simulations were conducted on the core components against key ferroptosis-related targets, glutathione peroxidase 4(GPX4), ACSL4, LPCAT3, and ALOX15, to assess binding stability. Spatial transcriptomics was utilized to directly observe, within the tissue spatial dimension, the distribution and expression patterns of key targets in atherosclerotic plaque regions of the aorta, thereby validating their association with pathological sites. An ApoE~(-/-)AS mouse model of AS induced by a high-fat diet was established. Mice were treated with different doses of Zuopi Pills to assess its effects on body weight, aortic pathology(hematoxylin-eosin staining), serum lipid profiles, inflammatory cytokines, and ferroptosis-related indices, as well as protein/mRNA expression in aortic tissue. Network pharmacology analysis identified 175 intersecting targets, with KEGG enrichment highlighting ferroptosis as a key pathway. Molecular docking showed favorable binding between core components and the targets, and molecular dynamics simulations confirmed the stability of these complexes. Spatial transcriptomics analysis visually demonstrated the specific high expression of GPX4, ACSL4, LPCAT3, and ALOX15 within the aortic plaque region, providing in situ evidence to support subsequent mechanistic studies. Animal experiments confirmed that Zuopi Pills significantly reduced body weight, improved serum lipid profiles, alleviated plaque formation, and inhibited inflammation in AS mice. Mechanistically, Zuopi Pills decreased Fe~(2+) and malondialdehyde(MDA) content while increasing superoxide dismutase(SOD) activity and glutathione(GSH) levels in aortic tissue. It downregulated the protein and mRNA expression of prostaglandin-endoperoxide synthase 2(PTGS2), transferrin receptor 1(TFR1), ACSL4, LPCAT3, and ALOX15, while concurrently upregulating GPX4 expression. In conclusion, Zuopi Pills exert a clear therapeutic effect on AS mice, and its mechanism is closely related to the regulation of the ACSL4/LPCAT3/ALOX15 signaling axis and the inhibition of ferroptosis in vascular cells.
PMID:42543366 | DOI:10.19540/j.cnki.cjcmm.20260305.705