Integrating serum pharmacochemistry and metabolomics to reveal the potential effective ingredients and mechanism of Huangqi Chifeng Tang intervening carotid atherosclerosis.

ETHNOPHARMACOLOGICAL RELEVANCE

As a representative herbal formula in traditional Chinese medicine (TCM), Huangqi ChiFeng Tang (HQCFT) is composed of Astragalus membranaceus (Fisch.) Bunge, Paeonia lactiflora Pall., and Saposhnikovia divaricata (Turcz.) Schischk. It has been clinically employed for managing carotid atherosclerosis (CAS) through its multi-component synergistic therapeutic effects. However, the material basis and mechanism of its pharmacological effects have not been systematically elucidated.

AIM OF THE STUDY

To systematically elucidate the pharmacological substance basis and mechanism of action of HQCFT intervention in CAS.

MATERIALS AND METHODS

Experiments were performed with high fat and high cholesterol diet (HFHCD) and carotid artery ligation induced CAS rats. Identification of chemical components of HQCFT and blood components after oral administration in rats using UPLC-Q-Exactive Orbitrap/MS technology. Non-targeted metabolomics was used to analyze the metabolite profile of rat serum after HQCFT intervention. In vivo anti-CAS effects of HQCFT assessed by H&E staining, lipid levels and blood mobility assays. ELISA was used to detect serum inflammatory factor levels. Immunofluorescence and Western blot were performed to detect protein levels of NLRP3 inflammasome and endothelial injury factor in carotid arteries.

RESULTS

A total of 169 components were identified by in vitro characterization of HQCFT chemical components, and 52 components were identified in serum samples, of which 18 were prototype components and 34 were metabolites. Non targeted serum metabolomics identified a total of 19 differential metabolites, mainly enriched in metabolic pathways such as arachidonic acid metabolism and ether lipid metabolism. HQCFT attenuated carotid artery pathological injury, ameliorated hepatic steatosis and dyslipidaemia, and inhibited the overexpression of serum inflammatory factors and NLRP3 inflammasome in CAS rats. HQCFT intervention notably inhibited the overexpression of serum inflammatory factors ICAM-1, VCAM-1, IL-1β, IL-6, IL-18, and CRP, as well as the protein levels of ICAM-1, VCAM-1, CD31, p-NF-κB/NF-κB, NLRP3, Caspase-1, and ASC.

CONCLUSIONS

Based on multi-omics analysis, this study elucidated the pharmacodynamic material basis of HQCFT and the regulation of serum metabolic profile in CAS rats. Further studies confirmed that HQCFT could effectively improve endothelial injury in CAS rats, and its potential mechanism was closely linked to its control of the NLRP3 inflammasome.