Study on the targets and mechanisms of 7-hydroxyethyl chrysin in prevention and treatment of high-altitude cerebral edema using TMT proteomics technology.

OBJECTIVES

To investigate the targets and mechanism of 7-hydroxyethyl chrysin (7-HEC) in prevention and treatment of high-altitude cerebral edema (HACE) in rats.

METHODS

Fifty-four male Wistar rats were randomly divided into normal control group, HACE model group, and 7-HEC-treated group (18 rats in each group). Except for the normal control group, rats in the other two groups were exposed to a hypobaric hypoxic chamber simulating 7000 m altitude for 72 h to establish the HACE model. The 7-HEC-treated group received intraperitoneal injection of 7-HEC (150 mg·kg¹·d¹) for 3 days prior to modeling, while the model group received equivalent saline. Tandem Mass Tag (TMT) proteomics technology was used to detect differentially expressed proteins (DEPs) with screening criteria set at a fold change >1.2 and <0.05. Western blotting was used to verify the expression levels of target proteins. Gene Ontology (GO) enrichment analysis, Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis, and protein-protein interaction (PPI) network analysis were performed.

RESULTS

Compared with the normal control group, 256 DEPs were identified in the HACE model group. Com-pared with the HACE model group, 87 DEPs were identified in the 7-HEC-treated group. Among them, 19 DEPs that were dysregulated in the HACE model group were restored after 7-HEC intervention, of which seven (HSPA4, Arhgap20, SERT, HACL1, CCDC43, POLR3A, and PCBD1) were confirmed by Western blotting. GO enrichment analysis of the DEPs between the HACE model and 7-HEC-treated groups revealed their involvement in 13 biological processes, five cellular components, and two molecular functions. KEGG pathway analysis indicated associations with the mRNA surveillance pathway, serotonergic synapse, RNA polymerase, protein processing in the endoplasmic reticulum, peroxisome, neuroactive ligand-receptor interaction, folate biosynthesis, cytokine-cytokine receptor interaction, and biosynthesis of cofactors. PPI network analysis demonstrated that HSPA4, POLR3A, and HACL1, which were validated by Western blotting, interacted with multiple signaling pathways and ranked among the top 20 hub proteins by degree value. Arhgap20, SERT (Slc6a4), and PCBD1 also exhibited interactions with several proteins, suggesting their potential as key regulatory proteins, whereas no interactions were detected for CCDC43.

CONCLUSIONS

This study applied TMT proteomics to identify seven potential therapeutic targets of 7-hydroxyethyl chrysin (7-HEC) for the prevention and treatment of HACE. These targets may participate in HACE prevention and treatment through multiple pathways, including maintaining cellular homeostasis, ameliorating oxidative stress, regulating energy metabolism, and reducing vascular permeability.