UPLC‐Q‐TOF‐MS‐Driven Systems Pharmacology Analysis of Dengzhan Shengmai Capsule Against Heart Failure: Integrating Serum/Tissue Distribution, Molecular Docking, and scRNA‐seq Evidence

Heart failure (HF) affects over 64 million individuals globally, and its prevalence continues to escalate, driven by population aging and the rising burden of metabolic disorders. Dengzhan Shengmai (DZSM) capsule, a classical Traditional Chinese Medicine formulation, has demonstrated therapeutic potential for HF. However, its bioactive constituents and underlying molecular mechanisms remain largely unexplored. We employed ultra-performance liquid chromatography-quadrupole time-of-flight mass spectrometry (UPLC-Q-TOF-MS) to characterize the metabolic profiles of DZSM in rat serum, cardiac, and hepatic tissues. Network pharmacology and bioinformatics approaches were subsequently applied to identify potential targets of these metabolites. Our integrated analytical framework encompassed protein-protein interaction (PPI) network construction, hub gene identification, Gene Ontology (GO) enrichment, Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis, miRNA-mRNA regulatory network analysis, core pathway evaluation, and gene-disease association mapping. Molecular docking, Western blot and RNA sequencing were performed to validate key bioactive components and their corresponding targets. We identified 67, 36, and 37 characteristic metabolites in serum, cardiac, and hepatic tissues, respectively. Critical bioactive compounds include caffeic acid, ferulic acid, quercetin-3-O-glucuronide, hyperoside, scutellarin, schizandrin A, and schizandrin B. The core therapeutic targets were identified as STAT3, CDK5, NOX4, and JAK2 in the chemokine signaling pathway. These biomarkers appear to influence HF treatment through modulation of CCL2-CCR2 axis, JAK-STAT, and MAPK signaling pathways. Molecular docking confirmed strong binding affinities between bioactive components and target proteins. Western blot results demonstrated that scutellarin dose-dependently inhibited the phosphorylation of JAK2 and STAT3 induced by oxygen-glucose deprivation/reoxygenation (OGD/R), with JAK2 and STAT3 total proteins remain almost unchanged. Single-cell RNA sequencing spatially mapped the expression patterns of key targets in myocardial tissue. Integrated serum/tissue pharmacochemistry and systems pharmacology elucidate DZSM's bioactive constituents and mechanistic foundations for HF intervention.