Angong Niuhuang Pill Attenuates Myocardial Infarction Through IL-17-Related Inflammatory Modulation and Mitochondrial Quality Control: Multi-Layer Analysis and Experimental Validation

Background: Acute myocardial infarction (AMI) remains the most lethal critical emergency worldwide. Although Angong Niuhuang Pill (ANP) is an established rescue medicine that has demonstrated outstanding therapeutic potential for cardiovascular diseases, its modern molecular mechanism has never been systematically elucidated because of its chemical complexity and unidentified targets. Methods: This study utilizes a multi-layer analytical pipeline of AI mining, network pharmacology, transcriptomics, and experimental confirmation. The components of ANP were comprehensively identified by UHPLC-Q Exactive Orbitrap HRMS. The TranSiGen algorithm was utilized to deeply mine the data and rank the components according to their relevance to AMI. The top 20 components were selected as prior weights and introduced into network pharmacology for analysis. Subsequently, a mouse model of AMI was established by ligating the left coronary artery. Cardiac function in the mice was evaluated by echocardiography and serum biochemical indicators. The pathological changes in the heart tissue were assessed by hematoxylin-eosin (H&E) and Masson staining. Cardiac transcriptome sequencing was performed, and pathway enrichment was analyzed by KEGG. The key pathways were verified by qPCR and immunofluorescence, achieving cross-validation between AI prediction and experimental findings. Results: The identification of ANP resulted in the detection of a total of 73 compounds, and the TranSiGen algorithm was employed to prioritize these compounds, yielding a ranked list of the top 20 candidates. Functional evaluation using echocardiography, serum biochemical markers, and histopathological examination demonstrated that ANP significantly ameliorated cardiac function in mice following myocardial infarction. Integration of network pharmacology and transcriptomic enrichment identified convergent axes of IL-17 signaling and mitochondrial quality control, which were subsequently experimentally validated as mechanisms by which ANP ameliorated cardiac injury. Experimental validation confirmed that ANP downregulated protein expression of IL-17A and TNF-α, normalized PINK1 and LC3-II/LC3-I marker profiles, with concomitant p62 reduction, thereby providing comprehensive molecular evidence at both transcriptional and translational levels to support the AI-driven predictions. Conclusions: This study identified IL-17 signaling and mitochondrial quality control as pathway axes associated with ANP-mediated cardioprotection against AMI, supported by AI-driven compound screening, transcriptome-network cross-validation, and experimental confirmation. This analytical framework may be adaptable to other complex TCM formulas for mechanism exploration and clinical translation.