α-Mangostin protects human brain microvascular endothelial cells from OGD/R-induced damage via regulation of the PDE4D-MAPK pathway

Ischemic stroke is a leading cause of neurological disability and death worldwide, and effective therapeutic agents targeting cerebral ischemia/reperfusion (I/R) injury remain urgently needed. α-Mangostin, a xanthone derivative isolated from mangosteen fruit, possesses neuroprotective activity. This study aimed to explore the neuroprotective effect of α-mangostin on oxygen-glucose deprivation and reperfusion (OGD/R)-stimulated human brain microvascular endothelial cells (hBMECs). Cell viability, Apoptosis, and Reactive Oxygen Species (ROS) content were evaluated by CCK-8 assay, flow cytometry, and ROS assay kit, respectively. Expression of phosphodiesterase 4D (PDE4D) was detected by qRT-PCR and western blot analysis. α-Mangostin targets were retrieved from SwissTargetPrediction and SEA databases and ischemic stroke-related targets were obtained from DisGeNet and GeneCards databases. A Venn diagram was used to identify overlapping targets. Targets of PDE4D in ischemic stroke were predicted using CTD database and subjected to KEGG pathway analysis. The changes in the mitogen-activated protein kinase (MAPK) pathway were examined by western blot analysis. Results showed that α-mangostin alleviated OGD/R-induced viability reduction and Apoptosis in hBMECs. Moreover, α-mangostin abolished OGD/R-triggered inflammatory response and ROS generation in hBMECs. PDE4D was identified as the only intersecting target of α-mangostin against ischemic stroke. α-Mangostin inhibited PDE4D expression and PDE4D overexpression reversed the effects of α-mangostin on OGD/R-induced injury in hBMECs. KEGG analysis showed that targets of PDE4D were enriched in MAPK pathway. α-Mangostin inactivated the MAPK pathway by downregulating PDE4D expression. In conclusion, α-mangostin attenuates OGD/R-induced injury in hBMECs through inhibiting the MAPK pathway by downregulating PDE4D expression. These findings provide an experimental basis for further research on α‑mangostin in cerebral ischemia/reperfusion injury.