OBJECTIVE: To systematically elucidate the cardioprotective mechanisms of the traditional Uyghur polyherbal formulation Yangxin Dawa Yimixike Mi Gao (YXDW, 养心达瓦) against myocardial ischemia/ reperfusion injury through integrated chemical analysis, bioinformatics prediction, and multilevel experimental validation.

METHODS: Using ultra-high-performance liquid chromatography coupled with quadrupole-Orbitrap high-resolution mass spectrometry (UHPLC-Q-Orbitrap-HRMS), 137 bioactive constituents were identified in YXDW. Potential targets associated with myocardial ischemia/reperfusion (I/R) injury were predicted through network pharmacology analysis and validated using Mendelian randomization (MR). Gene ontology and kyoto encyclopedia of genes and genomes enrichment analyses, together with molecular docking, were performed to identify the core regulatory pathways. An in vivo myocardial I/R model and an in vitro hypoxia/ reoxygenation (H/R) model were established in Sprague-Dawley rats and H9c2 cardiomyocytes, respectively. The therapeutic effects of YXDW were evaluated using histopathological examination, biochemical markers, including creatine kinase (CK), lactate dehydrogenase (LDH), superoxide dismutase, C-reactive protein, interleukin-6, and tumor necrosis factor-alpha, echocardiography, enzyme-linked immunosorbent assay, western blotting, and quantitative reverse transcription polymerase chain reaction.

RESULTS: UHPLC-Q-Orbitrap-HRMS fingerprinting demonstrated high batch-to-batch consistency, with similarity values greater than 0.87, and identified 23 compounds with high oral bioavailability (OB ≥ 56%). Network pharmacology and MR analyses identified 13 key targets associated with myocardial I/R injury, including C-X-C motif chemokine receptor 1 (CXCR1), mitogen-activated protein kinase kinase 1 (MAP2K1), and poly (ADP-ribose) polymerase 1 (PARP1), which may be involved in the regulation of apoptosis, oxidative stress, and inflammatory responses. In vivo experiments showed that YXDW administration alleviated I/R-induced myocardial injury, as evidenced by reduced serum CK and LDH levels, attenuated myocardial apoptosis, and improved left ventricular function. Furthermore, the in vivo and in vitro results showed that YXDW dose-dependently downregulated CXCR1, MAP2K1, and PARP1 expression at both the protein and messenger RNA (mRNA) levels in myocardial tissues and H/R-injured H9c2 cardiomyocytes.

CONCLUSION: YXDW exerts cardioprotective effects against myocardial I/R injury through multi-target modulation of inflammatory, oxidative, and apoptotic pathways. Our findings provide preliminary mechanistic support for the clinical relevance of YXDW in I/R therapy and offer a modern pharmacological basis for the rational use of traditional polyherbal formulations.