Protective effect of Hydroxysafflor Yellow A on cerebral ischemia reperfusion-injury by regulating GSK3β-mediated...

Ischemia-reperfusion (I/R) injury is accompanied by high mortality and morbidity. Unfortunately, there are few effective therapeutic medicines and strategies to enhance its outcome. Hydroxysafflor Yellow A (HSYA) exerts multiple biological activities and has potential protective effects against I/R injury in the brain, liver and heart. However, its underlying mechanism is still unclear. Here, we investigated whether HSYA modulates apoptosis and neuro-inflammation through the Glycogen synthase kinase-3β(GSK3β)-mediated pathway in a transient middle cerebral artery occlusion (MCAO) rat model and oxygen/glucose deprivation (OGD)-challenged primary neuronal cultures both in vivo and in vitro. Male Wistar rats were subjected to MCAO for 2 h, followed by 24 h of reperfusion. HSYA was administered 15 min after occlusion, SB216763 (GSK3β inhibitor) was injected to the left ventricle of the rat 6 h prior to MCAO. After 24 h of perfusion, apoptosis-associated protein and inflammatory markers were detected by western blotting. Meanwhile, terminal-deoxynucleotidyl transferase mediated nick end labeling(TUNEL) assay was used to evaluate the number of apoptotic cells in OGD-challenged neurons, cleaved caspase-3 were evaluated by Immunofluorescence (IF). Our data indicated that HSYA administration reduced infarct volume, decreased neurological deficit scores, elevated GSK3β phosphorylation and inhibited the activation of iNOS, NF-κB, and capase-3 in the penumbra of I/R rats. Moreover, blockade of GSK3β partly reversed the protective effect of HSYA on I/R by regulating NF-κB and caspase-3 both in vivo and in vitro. Collectively, we found that HSYA ameliorates I/R injury through its anti-inflammatory and anti-apoptotic effects via modulation of GSK-3β phosphorylation.