Neuroprotective efficacy of Curcumae Rhizoma against ischemic stroke via attenuation of COX-2-mediated neuroinflammation: a systematic study predicting thrombin inhibition and highlighting alexandrin and hederagenin.

Ischemic stroke is a major cause of mortality globally, but single-target therapies often fail due to the complex "ischemic cascade". This study investigated the neuroprotective effects and multitarget mechanisms of Curcumae Rhizoma (CR) using an integrated network pharmacology approach and in vivo validation. The active compounds of CR were screened through ADME filtering, and their potential targets were identified using the TCMSP, TTD, and UniProt databases. Molecular docking was used to validate the structural binding stability between CR ligands and core targets. In vivo efficacy was evaluated using a mouse model of middle cerebral artery occlusion, followed by histological analysis of infarct volume, neurological scores, and markers of reactive astrogliosis (GFAP) and neuroinflammation (COX-2). Network analysis identified 20 active compounds and 49 core targets, including F2 (thrombin) and PTGS2 (COX-2). Molecular docking revealed that alexandrin and hederagenin exhibited potent binding affinities for COX-2 and thrombin. In vivo, CR extract (CRex; 100 mg/kg) significantly reduced infarct volume and cerebral edema, while facilitating functional motor recovery. Mechanistically, CRex suppressed the overactivation of GFAP-positive reactive astrocytes (an intermediate filament marker of reactive astrogliosis) and downregulated COX-2 expression in the peri-infarct cortex. These findings demonstrate that CRex exerts synergistic neuroprotective effects primarily by attenuating COX-2-driven neuroinflammation, while network pharmacology predicts its secondary potential to interact with thrombin networks. By shifting the focus toward computationally predicted high-affinity candidates, such as alexandrin and hederagenin, our findings establish a valuable molecular hypothesis for CR as a promising multi-target therapeutics for ischemic stroke management.