Artesunate protects against ischemic stroke through THBS2/CD36/TLR4 pathway.

Abstract

Stroke remains a major global health challenge, causing irreversible neuronal damage with limited therapeutic options. Artesunate (ART), a derivative of artemisinin, has shown anti-inflammatory and neuroprotective properties, but its mechanisms in protecting against ischemic stroke are not fully elucidated. This study investigated the potential mechanisms of ART in mitigating experimental ischemic stroke using a transient middle cerebral artery occlusion (tMCAO) mouse model and cellular oxygen-glucose deprivation/reperfusion (OGD/R) models. The results demonstrated that ART significantly reduced infarct volume, improved neurological and motor functions, and suppressed neuronal apoptosis, oxidative stress, and inflammation in the tMCAO model. Transcriptomic sequencing and subsequent validation suggested that ART exerts its effects by regulating Thrombospondin 2 (THBS2) expression, with molecular docking and cellular thermal shift assay (CETSA) indicating a high affinity between ART and THBS2. In vitro experiments confirmed that ART attenuated OGD/R-induced damage and inflammation, while overexpression of THBS2 reversed these protective effects. Further mechanistic analysis revealed that ART protects against ischemic injury by inhibiting the THBS2/CD36/TLR4 signaling pathway and THBS2 overexpression abolished ART-induced neuroprotection. In conclusion, ART exhibits significant neuroprotective effects in experimental ischemic stroke through the downregulation of the THBS2/CD36/TLR4 pathway, highlighting its potential as a therapeutic agent.