The Study of the Protection Mechanism of Calycosin-7-O-β-d-Glucoside Against Oxygen–Glucose Deprivation/Reperfusion in HT22 Cells Based on Non-Targeted Metabolomics and Network Analysis

The cell non-targeted metabolomics technique was used to investigate the potential mechanism of Caly-cosin-7-O-β-d-glucoside (CAG) against cell oxygen–glucose deprivation/reperfusion (OGD/R). The OGD/R-injured HT22 cell model was constructed. The cells were divided into control, OGD/R, Edaravone (EDA), CAG-L, CAG-M, and CAG-H groups. The protective effect of CAG on OGD/R-injured nerve cells and its potential mechanism was investigated by detecting ROS levels, apoptosis rate, glutamic acid (Glu), γ-aminobutyric acid (GABA), nitric oxide (NO), and combining with cell non-targeted metabolomics. The results showed that after OGD/R, ROS levels, apoptosis rate, Glu and NO concentrations were significantly increased, while the concentrations of GABA were decreased considerably, which improved in a dose-dependent manner after CAG intervention. Cell non-targeted metabolomics results showed that CAG can dramatically improve the metabolomic characteristics of OGD/R-injured HT22 cells. Through bioinformatics analysis and molecular docking, it was found that purine metabolism may be an important pathway for CAG to treat OGD/R injury, and key proteins screened may be important targets for improving OGD/R injury. Therefore, CAG may protect OGD/R-injured HT22 cells by inhibiting apoptosis and oxidative stress, improving energy supply and the metabolomic characteristics of OGD/R-injured HT22 cells by regulating purine metabolism.