The potential value of quercetagetin in treating cerebral infarction by regulating miR-30d-5p/AKT1.

INTRODUCTION: Cerebral infarction (CI) represents a central nervous system vascular disorder characterized by high risks of disability and fatality. Quercetagetin (RG, 6-Hydroxyquercetin), a flavonoid isolated from Carthami Flos, has therapeutic potential against cerebral infarction. This research investigates its action mechanisms. METHODS: Human astrocytes (HAs) subjected to oxygen-glucose deprivation/reperfusion (OGD/R) and the middle cerebral artery occlusion-induced CI mice model were employed to investigate the protective effect of RG. By means of network pharmacology, AKT1 was identified as a potential CI-related target of RG in CI treatment. By predicting the ENCORI/starBase database and analyzing the GSE86291 dataset, miR-30d-5p was screened as a promising AKT1 regulator. Their regulatory relationship was verified through a dual-luciferase reporter experiment, and their expression was examined using the RT-qPCR or western blot technique. Furthermore, cell viability was detected using the CCK-8 assay, and cell apoptosis was evaluated via flow cytometry analysis. Additionally, Rotarod and Morris water maze tests were applied for the behavioral assessment of mice. RESULTS: RG exhibited a concentration-dependent protective effect by restoring cell viability and reducing apoptosis in OGD/R-treated HAs. MiRNA-30d-5p downregulated AKT1 expression by targeting its mRNA. Upregulation of miR-30d-5p exacerbated HA cell damage induced by OGD/R, and this damage was alleviated by AKT1 overexpression. miR-30d-5p agomir weakened RG's protective effect on OGD/R-treated HAs, and this suppression was reversed by the co-transfection of the AKT1 overexpression vector. RG downregulated the miR-30d-5p level while upregulating AKT1 expression in OGD/R-treated HAs and CI mice. Meanwhile, the motor coordination and learning/memory abilities of CI mice were enhanced. CONCLUSION: RG exerted a protective effect on HAs against OGD/R-induced impairment via the miR-30d-5p/AKT1 mechanism.