Anti‐diabetic mechanism and potential bioactive peptides of casein hydrolysates in STZ/HFD‐induced diabetic rats

Abstract BACKGROUND Casein hydrolysates have attracted much interest as anti‐diabetic food, but their hypoglycemic mechanism and biopeptides are not well understood. This study aimed to explore the anti‐diabetic mechanism and potential biopeptides of casein hydrolysates in streptozotocin/high‐fat‐diet‐induced diabetic rats and HepG2 cells. RESULTS Oral administration of casein hydrolysate prepared with papain–Flavourzyme combination (P‐FCH) decreased fasting blood glucose, improved oral glucose tolerance, and reduced HbA1c values in diabetic rats. P‐FCH was ineffective in alleviating insulin resistance (homeostasis model assessment and insulin sensitivity index) and enhancing hepatic insulin signaling transduction (phosphorylated Akt, hexokinase activity, and pyruvate kinase activity) in diabetic rats. However, P‐FCH significantly upregulated adenosine monophosphate‐activated protein kinase phosphorylation and glucose transporter‐2 expression, inhibited phosphoenolpyruvate carboxylase kinase activity, and elevated glycogen content in liver tissue of diabetic rats. Furthermore, P‐FCH increased glucose consumption independently in normal and insulin‐resistant HepG2 cells without the presence of insulin. The peptide composition of P‐FCH was characterized. The potential biopeptides in P‐FCH showed the sequence characteristic of a Val at the N‐terminal or a Pro at the P2 position, and the hypoglycemic activity of Val‐Pro‐Leu‐Gly (the most potential biopeptide in P‐FCH) was verified by oral glucose tolerance test in mice. CONCLUSION These results suggested that activation of the non‐insulin‐mediated AMPK pathway might be the determinant mechanism of P‐FCH on the hypoglycemic effect. The novel peptide Val‐Pro‐Leu‐Gly in P‐FCH was effective in reducing blood glucose levels when orally administered to mice. © 2023 Society of Chemical Industry.