Abstract |
Oxidants and inflammation are well-known to be common causes for numerous diseases in the human body. The use of natural protein hydrolysates has become increasingly popular for treating oxidative-associated and inflammatory diseases. Crocodylus siamensis hemoglobin hydrolysates (CHHs) were obtained by pepsin digestion at different incubation times (2, 4, 6 and 8h). CHH obtained by 6-h hydrolysis (6h-CHH) showed the strongest antioxidant activity against nitric oxide radicals, while the highest anti-inflammatory activity with respect to decreasing nitric oxide (NO) production was found for 2h-CHH. To evaluate the antioxidant and anti-inflammatory activity of individual peptide components, 2h-CHH and 6h-CHH were purified by semi-preparative HPLC. Peptide fraction P57 isolated from 6h-CHH was found to exhibit the highest nitric oxide radical inhibition activity (32.0%). Moreover, purification of 2h-CHH yielded peptide fraction P16, which displayed a high efficacy in decreasing NO production of macrophage RAW 264.7 cells (83.2%) and significantly reduced pro-inflammatory cytokines and inflammatory mediators interleukin-6 (IL-6), interleukin-1 beta (IL-1β) and prostaglandin-E2 (PGE2) production to about 2.0, 0.3 and 1.9 ng/mL, respectively. Active peptide sequences were identified as antioxidant KIYFPHF (KF7), anti-inflammatory SAFNPHEKQ (SQ9) and IIHNEKVQAHGKKVL (IL15). Accordingly, CHHs could potentially serve as an alternative source of antioxidant and anti-inflammatory agents. |