Exploring the Mechanisms of Polygonum orientale L. Against Myocardial Ischemia: An Integrated Analysis Using Ultra‐High‐Performance Liquid Chromatography–Quadrupole Exactive Orbitrap Mass Spectrometry, Network Pharmacology, and RNA Sequencing

ABSTRACT Polygonum orientale L. (PO) represents significant bioactivities in treating myocardial ischemia (MI); however, its underlying mechanisms remain unclear. This study aims to elucidate PO's potential mechanisms in MI using an integrated approach that combines UHPLC–Q‐Exactive Orbitrap HRMS, network pharmacology, and RNA‐sequencing (RNA‐seq). Initially, the chemical constituents of PO were identified using UHPLC–Q‐Exactive Orbitrap HRMS. Subsequently, network pharmacology, molecular docking, and RNA‐seq were employed to screen potential active components of PO targeting MI and predict their molecular mechanisms. Then, the molecular mechanisms were verified using western blotting and ELISA in MI mice. A total of 45 components were identified from PO, with 14 potential active compounds interacting with 204 MI‐related genes. The findings suggest that PO could alleviate heart damage. The RNA‐seq results indicated 244 potential targets regulated by PO. Integrating RNA‐seq and network pharmacology analyses revealed that the toll‐like receptor signaling pathway plays an important role, alongside the PI3K‐Akt. Notably, PO reduced the expression of TLR4 and TLR2 while increasing p‐Akt and p‐PI3K levels in MI mice, leading to decreased inflammatory cytokines and apoptosis‐related proteins. This study provides initial evidence that PO inhibits the toll‐like signaling pathway and activates PI3K–Akt signaling pathway to exert protective effects against MI.