Taoren Honghua Decoction alleviates atherosclerosis by inducing autophagy and inhibiting the PI3K-AKT signaling pathway to regulate cholesterol efflux and inflammatory responses.

BACKGROUND

Taoren Honghua Decoction (THD) is a traditional Chinese formula known for enhancing blood circulation and demonstrating clinical efficacy in the treatment of cardiovascular and cerebrovascular diseases. However, the primary active components and the underlying mechanisms by which THD exerts its therapeutic effects on atherosclerosis (AS) remain insufficiently characterized.

OBJECTIVE

This study aims to systematically validate the protective effects of THD on AS and elucidate its potential molecular mechanisms through an integrative approach involving network pharmacology, in vivo, and in vitro experiments.

METHODS

The main active ingredients and corresponding targets of all traditional Chinese medicines in THD were collected from the TCSMP and BATMAN-TCM databases. Potential targets of AS were identified using the OMIM, DrugBank, DisGeNET, and CTD databases, and AS microarray gene data were obtained from the GEO database. A drug active ingredient-target relationship network and a PPI network were constructed using Cytoscape 3.9.2 software. The molecular functions of the core targets were annotated through GO and KEGG enrichment analyses to further elucidate the potential molecular mechanisms of THD's anti-AS effects. The ApoE-/-mouse AS model was constructed through a high-fat diet (HFD), and RAW264.7 macrophage model was induced with ox-LDL to further validate the results of network pharmacology.

RESULTS

Network pharmacology analysis revealed that the main five active ingredients of THD include quercetin, apigenin, luteolin, kaempferol, and tanshinone IIA. Subsequently, by analyzing the intersection genes of the main active ingredient targets of THD and the AS targets, a total of ten core targets were identified: TP53, PPARG, JUN, AKT1, INS, IL6, SIRT1, TNF, ESR1, and STAT3. These are considered the core targets of THD in the treatment of AS. The GO and KEGG enrichment analysis results indicate that THD may exert anti-AS effects by regulating lipid metabolism and the PI3K-AKT signaling pathway. In vivo and in vitro experiments showed that THD reduced circulating lipid levels, decreased intraplaque lipid accumulation, and increased intraplaque collagen fiber content in HFD-induced ApoE-/- mice. Additionally, THD reduced ox-LDL-induced macrophage-derived foam cell formation, inhibited the expression of inflammatory factors IL-6 and TNF-α, and promoted the expression of cholesterol efflux regulatory proteins PPARγ, ABCA1, and ABCG1. Notably, the autophagy inhibitor 3-MA reversed these effects, confirming that THD's action involves autophagy activation, evidenced by increased LC3II/I and decreased p62 levels.

CONCLUSION

This study demonstrates that THD exerts significant anti-AS effects through the inhibition of the PI3K/AKT signaling pathway and the activation of autophagy, thereby promoting cholesterol efflux and mitigating inflammation. By integrating network pharmacology with experimental validation, these findings provide a comprehensive understanding of THD's mechanisms in treating AS and offer a solid theoretical basis for its potential clinical application.