Network Pharmacology-based Prediction and In Vivo Validation of Red Ginger and Turmeric Extract Combinations against Hypercoagulability in Isoproterenol-Induced Rats

Background: Myocardial infarction is a critical cardiovascular condition often preceded by a hypercoagulable state. Red ginger (Zingiber officinale var. rubrum, ZOR) and turmeric (Curcuma longa L., CL) are recognized for their anticoagulant properties, yet their multi-target pharmacological interactions remain poorly understood. This study aimed to evaluate the anticoagulant potential of ZOR and CL combinations using an integrated network pharmacology and in vivo approach. Methods: Network pharmacology analysis was employed to predict potential molecular targets of ZOR and CL bioactive compounds. For in vivo validation, rats were treated with single extracts (300 mg/kg bw) or combinations (ratios 1:1, 1:3, 3:1), followed by isoproterenol-induced hypercoagulability. Anticoagulant activity was assessed by measuring Prothrombin Time (PT) and activated Partial Thromboplastin Time (aPTT). Results: Network pharmacology identified 10 key hub proteins, including STAT3, AKT1, and MTOR, indicating a multi-target mechanism. In vivo, isoproterenol effectively induced hypercoagulability. While ZOR (300 mg/kg BW) showed the highest individual potency, all combinations significantly prolonged PT and aPTT (p < 0.05). Based on Chou-Talalay analysis, the interaction was dose-dependent; PT showed nearly additive to synergistic effects (CI: 0.86– 1.07), whereas aPTT exhibited antagonistic trends (CI: 1.06–1.50) at higher ZOR ratios. Consequently, combination effects remained stable without exceeding the most potent single dose. Conclusion: The ZOR and CL combination provides a consistent anticoagulant response through additive interactions, likely mediated by the modulation of the AKT/mTOR signaling axis and coagulation factors. This integration offers a promising, safer multi-target strategy for preventing thrombotic events in myocardial infarction.