Establishment of a modified high glucose OGD/R cell model and its application in the study of diabetic cerebral ischemic injury

To establish an optimized HT22 cell diabetic oxygen-glucose deprivation/reoxygenation (OGD/R) model. The modification addresses existing limitations in in vitro models of diabetic cerebral ischemia-reperfusion injury, such as inconsistent modeling parameters, poor simulation of pathological processes, and insufficient injury specificity. Furthermore, we sought to verify the model’s validity and demonstrate the technical advantages of this modified method. Media with glucose concentrations of 1 g/L (6.25 mM, representing normal blood glucose) and 4 g/L (25 mM, representing diabetic hyperglycemia) were prepared using sugar-free Minimum Essential Medium (MEM). HT22 cells then underwent 24 h of adaptive culture, followed by 8 h of hypoxia in sugar-free, serum-free basal medium. Direct reoxygenation was conducted for 6 h, without medium replacement. The data demonstrated exposure to OGD/R led to a significant difference in cell viability between the two groups. High-glucose treatment resulted in a typical diabetic cerebral ischemia-reperfusion injury phenotype, characterized by a pronounced decrease in medium pH, significant morphological damage, and increased rates of autophagy and apoptosis relative to normal glucose controls. Additionally, reactive oxygen species (ROS) and inflammatory factor contents were elevated under high glucose conditions. A modified HT22-cell diabetic OGD/R model was successfully established, providing a reliable in vitro framework for basic research on diabetic cerebral ischemia-reperfusion injury.