Low-Molecular-Weight Hydrolysate from Black Goat Extract Had Antioxidative and Anti-Inflammatory Effects in Macrophage Cells via Inhibition of MAPKs and NF-κB Pathways

Abstract

This study evaluated the antioxidative and anti-inflammatory effects of a low-molecular-weight hydrolysate derived from black goat extract using alcalase, ProteAX, and trypsin (APT6 < 3 kDa). The APT6 < 3 kDa exhibited high antioxidative activities with estimated contents of 41.24% for antioxidative amino acids and 49.55% for hydrophobic amino acids. APT6 < 3 kDa inhibited nitric oxide and prostaglandin E2 production in RAW 264.7 cells induced by lipopolysaccharide (p<0.05), which was accompanied by the inhibition of inducible nitric oxide synthase and cyclooxygenase-2 protein expression. Additionally, APT6 < 3 kDa downregulated the mRNA expression and production of TNF-α and IL-1β (p<0.05). Mechanistically, these anti-inflammatory effects of APT6 < 3 kDa were shown by downregulating the MAPK and NF-κB pathways in a dose-dependent manner. Overall, 43 peptides were isolated from a certain fraction (F5) of APT6 < 3 kDa, demonstrating high antioxidative and anti-inflammatory activities. These results suggest that identified peptides from APT6 < 3 kDa possess natural antioxidants and hold potential as anti-inflammatory agents.

Beef peptides mitigate skeletal muscle atrophy in C2C12 myotubes through protein degradation, protein synthesis, and the oxidative stress pathway

“…Each enzyme was allowed to act for 2 h, resulting in a total hydrolysis process duration of 6 h. The concentrations and following processes were developed by a preliminary study. 19 After enzymatic hydrolysis, the mixture was heated to 95 °C for 10 min to inactivate the enzymes. The hydrolysate was then placed into an ultrafiltration tube (Vivaspin 20, Sartorius, Goettingen, Germany) and centrifuged at 4000 g for 30 min (Combi R515, Hanil Co. Ltd, Daejeon, Korea), which facilitated the collection of peptide fractions with a molecular weight under 3 kDa, referred to as “beef peptides (BPs)”.…”

“…The amino acid sequence of the selected RP-HPLC fraction was analyzed using a mass spectrometer (Triple TOF®5600+ system, AB SCIEX, Framingham, MA, USA) connected to an HPLC (Ultimate 3000, Thermo Dinex) system, following a modified method from a previous study. 19 Separation was performed on a column C8 (Zorbax 300SB-C8 RP, Agilent Technologies) using mobile phase A (0.1% formic acid in water) and mobile phase B (0.1% formic acid in acetonitrile), maintaining a flow rate of 0.3 mL min −1 . Specific details of the linear gradient utilized are presented in Table S3 †.…”