DST-3, a novel cryptotanshinone derivate, attenuates glutamate excitotoxicity after ischemic stroke via CREB-Homer1...

Ischemic stroke remains a leading cause of death and disability worldwide. While cryptotanshinone (CTS) shows therapeutic promise, its clinical application is hindered by poor pharmacokinetic properties. This study investigated DST-3, a novel derivative of CTS for enhanced neuroprotective effect against ischemic stroke with improved drug-like properties.

We systematically evaluated DST-3 using oxygen-glucose deprivation/reoxygenation (OGD/R)-injured neurons and middle cerebral artery occlusion (MCAO) rat model. Comprehensive analyses included apoptosis assessment, oxidative stress markers, behavioral tests, infarct volume measurement, glutamate release, calcium imaging, mitochondrial function assessment. RNA-sequencing analysis was conducted to explore the action mechanism of DST-3. Molecular mechanism studies were assessed using immunofluorescence, immunohistochemistry, western blot, qPCR and gene silencing.

DST-3 demonstrated superior neuroprotection compared to CTS in both in vitro and in vivo models. In OGD/R-injured neurons, DST-3 significantly reduced apoptosis, oxidative stress, and mitochondrial dysfunction. In MCAO rats, DST-3 treatment markedly improved neurological deficits, reduced infarct volume, and decreased neuronal apoptosis. Mechanistically, DST-3 inhibited glutamate excitotoxicity by maintaining calcium homeostasis and preserving mitochondrial function. RNA-sequencing revealed that Homer 1 as the crucial gene regulated by DST-3. Further investigation demonstrated that DST-3 upregulated Homer1 through CREB phosphorylation. CREB-Homer1 axis was identified as a potential protective target of DST-3.

DST-3 demonstrates enhanced neuroprotective efficacy through modulation of glutamate excitotoxicity and mitochondrial function, with the CREB-Homer1 signaling axis identified as a key underlying mechanism, establishing DST-3 as a promising therapeutic candidate for ischemic stroke treatment.