Physical and chemical properties and anti-apoptotic effects on C2C12 cells of hydrolyzed tuna (Katsuwonus pelamis) blood powder

Abstract

This study aimed to develop hydrolyzed tuna blood powders (HTBP) with potential anti-apoptotic and muscle recovery properties. HTBP was produced alcalase, trypsin, and pepsin at concentrations of 0.5%, 1.0%, and 1.5%. The physical and chemical properties of HTBP were evaluated. The results showed no significant difference in yield (p > 0.05), with a range of 7.08% ± 0.78% to 9.26 % ± 1.74%. HTBP produced with alcalase at all concentrations yielded a higher amount of protein (76.86% ± 0.5%–79.1% ± 1.10%) compared to trypsin and pepsin. Glutamic acid, cysteine, and aspartic acid were identified as the main amino acids in HTBP. The water solubility index of all HTBP was high (>90%). Antioxidant values increased with higher protein content. Hydrolyzed tuna blood with 0.5% alcalase (HTBPA) and 0.5% trypsin were selected for further investigation of their biological activities, assessed using C2C12 muscle cells under oxidative stress conditions induced by hydrogen peroxide (H₂O₂). In H₂O₂-induced C2C12 cells, HTBPA and HTBPT reduced nitric oxide (NO) secretion and intracellular reactive oxygen species (ROS) levels. Furthermore, HTBPA and HTBPT significantly decreased the percentage of apoptotic cells in these H₂O₂-induced C2C12 cells. They also inhibited H₂O₂-induced apoptosis by modulating apoptosis-related proteins, including NF-κB, Bax, and caspase-3. Additionally, HTBPA and HTBPT suppressed the phosphorylation levels of key signaling proteins such as Akt, ERK1/2, and p38, thereby contributing to enhanced cell survival. These findings suggest that hydrolyzing tuna blood, waste of the canned tuna industry, can create a valuable ingredient for promoting muscle health.