Deciphering the effective components of a TCM formula for atherosclerosis by three-dimensional pattern recognition of exogenous components correlated with endogenous metabolites.

Ethnopharmacological Relevance: The deciphering of effective components is crucial for understanding the role they play and how they function in traditional Chinese medicine (TCM) formulae. However, this remains a significant challenge for these complex systems with multiple components, targets, and pathways, despite their therapeutic benefits.

Aim Of The Study: Three-dimensional pattern recognition of exogenous components correlated with endogenous metabolites was proposed to discover the effective components of Gualou-Xiebai-Banxia decoction (GXB), a famous classical TCM formula for effective improvement of atherosclerosis (AS).

Materials And Methods: The potential effective exogenous components were determined by three-dimensional pattern recognition of abundance, bioavailability and AS-related activity. The efficacy of GXB in attenuating AS was evaluated using an Apolipoprotein E-deficient (ApoE) mice model subjected to a high-fat diet regimen. Plasma metabolomics was developed to dig out GXB efficacy-related endogenous metabolites. Next, the potential effective exogenous components and GXB efficacy-related endogenous metabolites were combined with AS targets to develop correlation analysis, so as to explore candidate effective components and potential mechanisms of GXB. Further, the effective components were validated by oxidized low-density lipoprotein-induced RAW 264.7 macrophages.

Results: A total of 30 potential effective exogenous components in GXB were ascertained by three-dimensional pattern recognition after conducting Technique for Order of Preference by Similarity to Ideal Solution (TOPSIS) analysis. GXB demonstrated a significant ameliorative effect on atherosclerotic symptoms in ApoE mice under a high-fat diet, as evidenced by decreasing serum lipid levels, atherosclerotic plaques (aorta and aortic root) and IL-6 content. Subsequently, metabolomics results revealed that it was associated with the regulation of endogenous metabolites, including organic acids, amino acid, fatty acids and glycerophospholipid. Next, the correlation analysis was constructed with AS targets by the network of "potential effective exogenous components-AS targets-endogenous metabolites", tentatively inferring that 18 exogenous components were candidate effective components, and lipid metabolism was the major regulation pathway of GXB. Furthermore, GXB suppressed lipid accumulation in vivo/vitro through increasing expressions of PPAGγ, ABCA1, ABCG1, and SR-B1 related to cholesterol efflux. Cucurbitacin B and 5 (6)-ene-macrostemonoside B were demonstrated as the effective components with inhibitory activity on foam cell formation and lipid accumulation.

Conclusion: Three-dimensional pattern recognition of exogenous components correlated with endogenous metabolites was proposed and effectively utilized to demystify the effective components of GXB in AS prevention. This strategy also provided a reference for the related studies of other classical TCM formulae.