Pharmacological Effects and Candidate Bioactive Constituents of Huanglian Wendan Decoction in Experimental Metabolic Syndrome: Multi-Omics Analysis of Gut–Liver Metabolism and NLRP3 Signaling

Abstract

Background Chronic low-grade inflammation and gut microbiota dysbiosis contribute to metabolic syndrome (MS). Huanglian Wendan Decoction (HLWDD) is a defined multi-herbal formulation used in Chinese medicine, but its effects on gut microbiota-associated metabolism and NLRP3 inflammasome signaling remain unclear. Objective This study evaluated the pharmacological effects and candidate absorbed bioactive constituents of HLWDD in experimental MS. Methods A high-fat, high-sugar diet-induced rat model of MS was assessed using metabolic, biochemical, histological, molecular, and multi-omics endpoints. Gut metagenomics, untargeted metabolomics of liver and colonic contents, liver transcriptomics, and UPLC-HRMS/MS analysis of serum-absorbed constituents were integrated with network pharmacology and molecular docking. Baicalin was further evaluated in PA-BSA-treated HepG2 cells. Results HLWDD ameliorated obesity, dyslipidemia, hepatic steatosis, and systemic inflammation and attenuated NLRP3 inflammasome-related signaling. Multi-omics analysis identified nine shared differential metabolites and a strong positive association between Faecalibacterium and butyrate. Thirty-two serum-absorbed constituents were tentatively identified. Baicalin showed favorable predicted interactions with NLRP3 (− 5.82 kcal/mol) and CASP1 ( − 5.41 kcal/mol) and concentration-dependently suppressed NLRP3 inflammasome-related signaling in HepG2 cells; the highest concentration increased intracellular butyrate. Conclusion HLWDD ameliorated metabolic and inflammatory abnormalities in experimental MS in association with restoration of gut–liver metabolic homeostasis and attenuation of NLRP3 inflammasome signaling. Baicalin emerged as a candidate absorbed bioactive constituent.