Investigation of the Mechanism of Total Flavonoids From Chuju in Mitigating Cerebral Ischemia–Reperfusion Injury in Rats Using Untargeted Metabolomics

ABSTRACT Chuju ( Dendranthema morifolium (Ramat.) Tzvel. cv.), a traditional Chinese medicinal herb with both medicinal and edible properties. It is widely used in tea beverages and functional foods due to its rich flavonoid and polysaccharide content. Our previous studies demonstrated that the total flavonoids of Chuju (TFCJ) alleviate cerebral ischemia–reperfusion injury (CIRI) in rats. However, the potential metabolic pathways regulated by TFCJ remain unclear, and its therapeutic targets require further investigation. This study aims to elucidate the neuroprotective mechanisms of TFCJ. LC–MS analysis identified 42 compounds within TFCJ. Animal experiments showed that TFCJ significantly reduced cerebral infarct volume, elevated SOD and GSH‐Px levels, and decreased MDA levels in the brain. Non‐targeted metabolomic analysis suggested that TFCJ exerts its protective effects by modulating the tricarboxylic acid (TCA) cycle, glycolysis, pentose phosphate pathway (PPP), and aspartate–glutamate metabolism. Pathway analysis revealed that the 4EBP1/eIF4E signaling pathway may mediate the protective effects of TFCJ. In conclusion, this study analyzed the main components of TFCJ and provided evidence suggesting that the 4EBP1/eIF4E signaling pathway might serve as a potential mediator of its neuroprotective effects against CIRI. These findings provide a theoretical basis for the application of TFCJ as both a functional food and a medicinal agent.