Hypoglycemic effects and mechanisms of animal-derived blood peptides in C2C12 myotubes and alloxan-induced diabetic mice

This study aimed to elucidate the hypoglycemic effects and mechanisms of animal-derived blood peptide protein powder (BPPP). BPPP dose-dependently increased glucose uptake in C2C12 myotubes (100, 200, 400 μg/mL) without compromising cell viability. Mechanistically, BPPP activated the insulin signaling pathway and promoted GLUT4 membrane translocation. Four-week BPPP gavage ameliorated diabetic symptoms in alloxan-induced mice, significantly lowering fasting glucose, improving glucose tolerance, and attenuating tissue damage. Moreover, BPPP enhanced hepatic antioxidant capacity and improved dyslipidemia in diabetic mice. Consistent with the in vitro experiments, BPPP upregulated the insulin signaling pathway and GLUT4 expression in skeletal muscle. Additionally, BPPP suppressed hepatic gluconeogenesis (PEPCK, G6Pase) while enhancing glycogen storage. LC-MS/MS analysis of BPPP-G1 and BPPP-G2 fractions identified three non-toxic bioactive peptides (YPWTQ, VDPENFRLL, RYVDPENFRLL), which were validated by Peptide Ranker and molecular docking. These findings provide a theoretical basis for BPPP application as a functional food ingredient or therapeutic agent in diabetes management. • BPPP enhanced glucose uptake in C2C12 myotubes and relieved diabetes in mice. • BPPP activated the IRS1/PI3K/AKT/GSK3β signaling pathway in vitro and in vivo . • Hypoglycemic effect of BPPP is mediated through upregulation of GLUT4 expression and ranslocation. • Hypoglycemic peptides were identified from BPPP using LC-MS/MS and bioinformatics analyses.