Objective Based on Astral-data-independent acquisition (DIA) proteomics technology, to screen the potential targets of agiophyllum oligo saccharides (AOS) to improve type 2 diabetes mellitus (T2DM) related liver injury, and explore its mechanism.

Methods Twelve db/db mice were randomly divided into a model group, a low-dose AOS group (LAOS, 375 mg/kg), and a high-dose AOS group (HAOS, 750 mg/kg), with 4 mice in each group. Additionally, 4 db/m mice were set as a blank control group. After 8 weeks of intervention, liver tissues were collected for proteomic analysis, and the mechanism of AOS was analyzed using methods such as Venn diagram, KEGG pathway enrichment, gene ontology (GO) annotation, protein-protein interaction (PPI) network, and gene set enrichment analysis (GSEA).

Results  180 differentially expressed proteins were identified in LAOS group. GO analysis showed that they were involved in breakdown and metabolism of cellular amino acid. KEGG suggested that they were related to the pathways of butyric acid metabolism and degradation of valine, leucine and isoleucine. 280 differentially expressed proteins were identified in HAOS group. They were significantly enriched in processes of small molecule metabolism, organic acid metabolism, ketone acid metabolism and carboxylic acid metabolism. KEGG analysis showed that they exerted their effects through regulating the pathways such as metabolic pathways, propionic acid metabolism, carbon metabolism, oxidative phosphorylation, and degradation of valine, leucine and isoleucine. PPI network screened 10 core target proteins including ACSS1, ECHDC1, ECHS1 and so on. GSEA showed that the role of LAOS was related to activation of  IFN-γ signaling and hypoxic pathways, while HAOS was involved in pathways of epithelial mesenchymal transition, hypoxia, unfolded protein response, muscle cell generation and protein secretion.

Conclusion  AOS might improve liver injury in diabetes by regulating branched chain amino acid metabolism, short chain fatty acid metabolism, pyruvate metabolism and oxidative phosphorylation. Targets such as ACSS1, ECHDC1, ECHS1 etc. were its key action nodes.

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