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Antioxidants (Basel)
2024 Aug 14;138:. doi: 10.3390/antiox13080988.
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Sea Cucumber Viscera Processed by Protease Hydrolysis Combined with Cordyceps militaris Fermentation Protect Caco-2 Cells against Oxidative Damage via Enhancing Antioxidant Capacity, Activating Nrf2/HO-1 Pathway and Improving Cell Metabolism.
Mi R
,
Fu Z
,
Jiang J
,
Gao S
,
Guan X
,
Wang X
,
Zhou Z
.
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Excessive reactive oxygen species (ROS) may lead to oxidative damage and metabolic disorder. The pathogenesis of human bowel inflammation is closely related to oxidative damage of intestinal epithelial cells caused by ROS. This study aimed to explore the high-value utilization of the byproducts of sea cucumber in antioxidant food for colitis prevention. The technology of protease hydrolysis combined with Cordyceps militaris fermentation was used to obtain fermented sea cucumber viscera protease hydrolysates (FSVHs). The results revealed that FSVH could enhance antioxidant capacity and alleviate oxidative damage and apoptosis by activating the Nrf2/HO-1 pathway and triggering the self-protection immune mechanisms. Moreover, the FSVH supplementation could upregulate antioxidant-related metabolic pathways of Caco-2 cells such as glutathione metabolism, confirming the enhanced antioxidant capacity of damaged cells. In summary, FSVH could exert protective effects on Caco-2 cells in response to oxidative damage, providing a promising prospect for sea cucumber resource utilization and colitis prevention.
2021RT08 Innovation and Entrepreneurship Program for High-level Talent of Dalian, 2021-84 Liaoning Marine Economic Development Project, 2021JH2/10200042 Science and Technology Projects in Liaoning Province, CARS-18 Earmarked Fund for Modern Agro-industry Technology Research System
Figure 1. Effects of SVH (A) and FSVH (B) on cell viability of Caco-2 cells. Results were mean ± SD for three experiments. ** p < 0.01.
Figure 2. Protective effects of SVH (A) and FSVH (B) on H2O2-induced oxidative damage of Caco-2 cells. Results were mean ± SD for three experiments. * p < 0.05, ** p < 0.01.
Figure 3. Effects of SVH and FSVH on cell cycle and apoptosis in H2O2-treated Caco-2 cells. Cell cycle analysis of Caco-2 cells (A,C). The apoptosis ratio of Caco-2 cells (B,D). Results were mean ± SD for three experiments. ** p < 0.01.
Figure 4. Effects of SVH and FSVH on the mitochondrial membrane potential, the ROS levels and the antioxidant enzyme activities in H2O2-treated Caco-2 cells. The mitochondrial membrane potential of Caco-2 cells (A,B). The ROS production in Caco-2 cells (C,D). The levels of MDA (E). The levels of SOD (F). The levels of CAT (G). The levels of T-AOC (H). Results were mean ± SD for three experiments. ** p < 0.01.
Figure 5. Effects of SVH and FSVH on the mRNA expression levels of HO−1 and NQO−1 in H2O2-treated Caco-2 cells. The mRNA levels of HO−1 (A) and NQO−1 (B). Results were mean ± SD for three experiments. ** p < 0.01.
Figure 6. Effects of SVH and FSVH on the protein expression levels of HO−1, NQO−1 and NRF2 in H2O2-treated Caco-2 cells. The protein expression levels of HO−1 (A,B), NQO−1 (A,C), cytoplasmic NRF2 (A,D) and nuclear NRF2 (A,E). Results were mean ± SD for three individual experiments. ** p < 0.01.
Figure 7. The PCA, PLS-DA, replacement verification plot and OPLS-DA of the control, H2O2, SVH and FSVH groups. (A) PCA score plot in negative ion mode (R2X = 0.492, Q2 = 0.0639). (B) PLS-DA score plot in negative ion mode (R2X = 0.528, R2Y = 0.919, Q2 = 0.721). (C) Replacement verification plot in negative ion mode; intercept of the regression Q2 = −0.271272. (D) OPLS-DA score graph in negative ion mode. (E) PCA score plot in positive ion mode (R2X = 0.508, Q2 = 0.0668). (F) PLS-DA score plot in positive ion mode (R2X = 0.552, R2Y = 0.884, Q2 = 0.632). (G) Replacement verification plot in positive ion mode; intercept of the regression Q2 = −0.26698. (H) OPLS-DA score graph in positive ion mode.
Figure 8. Heat map of differential metabolites in negative ion mode.
Figure 9. KEGG pathway analysis of SVH (A) and FSVH (B) groups. Each dot indicates one metabolic pathway; the size and shade of points represent the correlation with the influence of the metabolic pathway.
Figure 10. Schematic diagram of protective mechanism of FSVH supplementation against H2O2-induced oxidative damage of Caco-2 cells.
