Objective To systematically identify traditional Chinese medicine (TCM) compound formulas with anti-atherosclerosis (AS) potential using a screening platform based on multi-strain apoeb gene knockout zebrafish models for anti-atherosclerosis TCM compounds

Methods Screening was conducted using a formula dictionary database, and compound formulas such as Dan Shen Fang, Shang Ni San, Jin Lingzi San, and Li Qi Hua Liang Tang were selected from 933 blood-activating and stasis-resolving formulas. Five strains of apoeb^－/－ zebrafish–AB, Tg(mpeg1:EGFP), Tg(lyzc:dsRed), Tg(gata1a:mCherry), and Tg(flk1:EGFP)—were fed a high-fat diet for 2 days, after which the AS model was established. DMSO (solvent control), rosuvastatin (western medicine positive control), compound Dan Shen Fang (TCM positive control), Shang Ni San, Jin Lingzi San, and Li Qi Hua Liang Tang were administered by bathing for 24 hours. Oil red O staining was used to detect lipid content, oxidative stress indicators, blood flow velocity, vascular endothelial cell damage, and the expression of inflammatory factors in zebrafish. The behavior patterns of inflammatory cells in zebrafish were observed, and the anti-AS efficacy of the Li Qi Hua Liang formulas was comprehensively evaluated.

Results  Compared with the AS model group, compound Dan Shen Fang and Shang Ni San improved lipid accumulation in the vascular lumen, reduced total cholesterol (TC), triglycerides (TG), and low-density lipoprotein cholesterol (LDL-C) levels, decreased oxidative stress response, inhibited the recruitment of neutrophils and macrophages, repaired vascular endothelial damage, improved blood flow velocity, and reduced the expression of inflammatory factors, thereby alleviating AS, with statistically significant differences. Jin Lingzi San and Li Qi Hua Liang Tang had limited effects on AS.

Conclusion This platform provides an efficient and targeted tool for the screening and research evaluation of anti-atherosclerosis TCM compound formulas, accelerates the research progress of anti-atherosclerosis TCM compound formulas, and lays a foundation for subsequent drug development and clinical application.