Integrating UPLC-Q-TOF-MS/MS, feature-based molecular networking, network pharmacology, and molecular docking to investigate the mechanism of Persicae Ramulus against myocardial ischemia

Ethnopharmacological relevance: Myocardial ischemia (MI) is a central pathophysiological driver of Cardiovascular Disease (CVD). It is the acute manifestation of coronary artery disease and angina pectoris, and precipitates life-threatening complications including myocardial infarction and heart failure. The fundamental defect is an imbalance between coronary oxygen supply and myocardial oxygen demand, which precipitates energetic failure and contractile dysfunction. Given its rising incidence and mortality, the development of effective preventive and therapeutic strategies has become a priority in contemporary cardiology research. Preliminary data indicate that Persicae Ramulus (PR) confers cardioprotection against ischemia-driven cardiovascular injury, yet its molecular mechanisms and targets remain elusive.

Aims of study: This study sought to systematically identify the chemical constituents of PR by integrating ultra-performance liquid chromatography-quadrupole time-of-flight tandem mass spectrometry (UPLC-Q-TOF-MS/MS) with feature-based molecular networking (FBMN). Network pharmacology and molecular docking were subsequently employed to delineate the bioactive constituents and potential mechanisms underlying the therapeutic efficacy of PR against MI.

Materials and methods: Liquid chromatography-tandem mass spectrometry (LC-MS/MS) was employed to comprehensively profile the chemical constituents of PR. Network pharmacology was subsequently used to screen bioactive ingredients and identify MI-related targets. Functional enrichment analysis was performed with the DAVID database and additional bioinformatics platforms. AutoDock Vina and PyMOL were then used for molecular docking and to visualize the binding modes of the key compounds. All predicted mechanisms were validated in both in vivo and in vitro models.

Results: Using a diagnostic-ion-guided and FBMN strategy, 141 constituents were tentatively identified in negative-ion mode, comprising 61 Flavonoids, 29 Phenylpropanoids, 15 organic acids, 14 glycosides, 12 fatty acids, 3 Triterpenes, and 7 miscellaneous compounds; 356 precursor ions (59 clusters with ≥2 nodes and 84 singletons) were generated. Network pharmacology indicated that 107 active ingredients potentially act on 443 MI-related targets. After visual filtering, 79 core targets (e.g., DNM1L, SIRT1, NFE2L2) and 89 core compounds were retained. KEGG enrichment highlighted Apoptosis, calcium signaling, and Toll-like Receptor pathways, among Others. Docking revealed 40 compounds with strong affinities (binding energy <-8.0 kcal mol^-1) toward SIRT1, PGC-1α, and DRP1. Experimentally, PR significantly reduced circulating levels of CRP, BNP, CK, and LDH, restored myocardial ATP content, preserved tissue architecture, and attenuated fibrosis. Mechanistically, PR up-regulated SIRT1/PGC-1α signaling and modulated DRP1 phosphorylation at Ser637 and Ser616 (p < 0.05, 0.01).

Conclusion: The bioactive constituents of PR confer cardioprotective effects via the SIRT1/PGC-1α axis. These findings establish a rational basis for future dissection of its chemical entities, pharmacodynamic actions, and translational potential in PR.

Feature-based molecular networking; Myocardial ischemia; Persicae ramulus; SIRT1/PGC-1α signaling pathway.