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Anti-inflammatory effects of a casein hydrolysate and its peptide-enriched fractions on TNFα-challenged Caco-2 cells and LPS-challenged porcine colonic explants.

Mukhopadhya A, Noronha N, Bahar B, Ryan MT, Murray BA, Kelly PM, O'Loughlin IB, O'Doherty JV, Sweeney T - Food Sci Nutr (2014)

Bottom Line: Bioactive milk peptides are reported to illicit a range of physiological benefits and have been proposed as potential functional food ingredients.While dexamethasone reduced the IL-8 concentration by 41.6%, the 1 kDaR and 1 kDaP fractions reduced IL-8 by 68.7% and 66.1%, respectively, relative to TNFα-stimulated Caco-2 cells (P < 0.05).In the ex vivo system, only the 1 kDaR fraction elicited a decrease inIL1-α,IL1-β,IL-8,TGF-β andIL-10 expression (P < 0.05).

View Article: PubMed Central - PubMed

Affiliation: School of Veterinary Medicine, UCD Dublin, Ireland ; Food for Health Ireland (FHI), UCD Belfield, Dublin, Ireland.

ABSTRACT
Bioactive milk peptides are reported to illicit a range of physiological benefits and have been proposed as potential functional food ingredients. The objective of this study was to characterize the anti-inflammatory properties of sodium caseinate (NaCAS), its enzyme hydrolysate (EH) and peptide-enriched fractions (5 kDa retentate [R], 1 kDaR and 1 kDa permeate [P]), both in vitro using a Caco-2 cell line, and also ex vivo using a porcine colonic tissue explant system. Caco-2 cells were stimulated with tumour necrosis factor alpha (TNFα) and co-treated with casein hydrolysates for 24 h. Following this, interleukin (IL)-8 concentrations in the supernatant were measured using enzyme-linked immunosorbent assay. Porcine colonic tissue was stimulated with lipopolysaccharide and co-treated with casein hydrolysates for 3 h. The expression of a panel of inflammatory cytokines was measured using qPCR. While dexamethasone reduced the IL-8 concentration by 41.6%, the 1 kDaR and 1 kDaP fractions reduced IL-8 by 68.7% and 66.1%, respectively, relative to TNFα-stimulated Caco-2 cells (P < 0.05). In the ex vivo system, only the 1 kDaR fraction elicited a decrease inIL1-α,IL1-β,IL-8,TGF-β andIL-10 expression (P < 0.05). This study provides evidence that the bioactive peptides present in the 1 kDaR fraction of the NaCAS hydrolysate possess anti-inflammatory properties in vitro and ex vivo. Further in vivo analysis of the anti-inflammatory properties of the 1 kDaR is proposed.

No MeSH data available.


Related in: MedlinePlus

Effect of NaCAS, EH, 5 kDaR, 1 kDaR or 1 kDaP on relative quantity (RQ) of a selected panel of cytokine genes stimulated by LPS challenge for 3 h in porcine colonic tissue explant. After challenge with LPS and treatment with milk hydrolysates, total RNA was extracted, cDNA synthesised and RT-qPCR was performed to quantify the selected panel of genes. RQ values of treatments with NaCAS, EH, 5 kDaR, 1 kDaR or 1 kDaP compared to LPS challenged control, error bars indicate SE. *P < 0.05, **P < 0.01.
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fig03: Effect of NaCAS, EH, 5 kDaR, 1 kDaR or 1 kDaP on relative quantity (RQ) of a selected panel of cytokine genes stimulated by LPS challenge for 3 h in porcine colonic tissue explant. After challenge with LPS and treatment with milk hydrolysates, total RNA was extracted, cDNA synthesised and RT-qPCR was performed to quantify the selected panel of genes. RQ values of treatments with NaCAS, EH, 5 kDaR, 1 kDaR or 1 kDaP compared to LPS challenged control, error bars indicate SE. *P < 0.05, **P < 0.01.

Mentions: The effect of NaCAS, NaCAS EH, retentates, and permeates on gene expression in the LPS-stimulated tissue is presented in Figure3. Co-treatment of colonic tissues with hydrolysates EH, 5 kDaR and 1 kDaR was associated with a downregulation ofIL1-α (0.086, 0.021, 0.028 vs. 0.424 ± 0.083,P < 0.05) andTNFα (0.101, 0.123, 0.128 vs. 0.234 ± 0.034,P < 0.05) expression relative to LPS-challenged tissues. Similarly, co-treatment with 5 kDaR and 1 kDaR hydrolysates was associated with a downregulation ofIL-1β (0.151, 0.096 vs. 0.442 ± 0.093,P < 0.05) andIL-8 (0.033, 0.068 vs. 0.212 ± 0.050,P < 0.05) expression relative to LPS-challenged tissues.


Anti-inflammatory effects of a casein hydrolysate and its peptide-enriched fractions on TNFα-challenged Caco-2 cells and LPS-challenged porcine colonic explants.

Mukhopadhya A, Noronha N, Bahar B, Ryan MT, Murray BA, Kelly PM, O'Loughlin IB, O'Doherty JV, Sweeney T - Food Sci Nutr (2014)

Effect of NaCAS, EH, 5 kDaR, 1 kDaR or 1 kDaP on relative quantity (RQ) of a selected panel of cytokine genes stimulated by LPS challenge for 3 h in porcine colonic tissue explant. After challenge with LPS and treatment with milk hydrolysates, total RNA was extracted, cDNA synthesised and RT-qPCR was performed to quantify the selected panel of genes. RQ values of treatments with NaCAS, EH, 5 kDaR, 1 kDaR or 1 kDaP compared to LPS challenged control, error bars indicate SE. *P < 0.05, **P < 0.01.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4256577&req=5

fig03: Effect of NaCAS, EH, 5 kDaR, 1 kDaR or 1 kDaP on relative quantity (RQ) of a selected panel of cytokine genes stimulated by LPS challenge for 3 h in porcine colonic tissue explant. After challenge with LPS and treatment with milk hydrolysates, total RNA was extracted, cDNA synthesised and RT-qPCR was performed to quantify the selected panel of genes. RQ values of treatments with NaCAS, EH, 5 kDaR, 1 kDaR or 1 kDaP compared to LPS challenged control, error bars indicate SE. *P < 0.05, **P < 0.01.
Mentions: The effect of NaCAS, NaCAS EH, retentates, and permeates on gene expression in the LPS-stimulated tissue is presented in Figure3. Co-treatment of colonic tissues with hydrolysates EH, 5 kDaR and 1 kDaR was associated with a downregulation ofIL1-α (0.086, 0.021, 0.028 vs. 0.424 ± 0.083,P < 0.05) andTNFα (0.101, 0.123, 0.128 vs. 0.234 ± 0.034,P < 0.05) expression relative to LPS-challenged tissues. Similarly, co-treatment with 5 kDaR and 1 kDaR hydrolysates was associated with a downregulation ofIL-1β (0.151, 0.096 vs. 0.442 ± 0.093,P < 0.05) andIL-8 (0.033, 0.068 vs. 0.212 ± 0.050,P < 0.05) expression relative to LPS-challenged tissues.

Bottom Line: Bioactive milk peptides are reported to illicit a range of physiological benefits and have been proposed as potential functional food ingredients.While dexamethasone reduced the IL-8 concentration by 41.6%, the 1 kDaR and 1 kDaP fractions reduced IL-8 by 68.7% and 66.1%, respectively, relative to TNFα-stimulated Caco-2 cells (P < 0.05).In the ex vivo system, only the 1 kDaR fraction elicited a decrease inIL1-α,IL1-β,IL-8,TGF-β andIL-10 expression (P < 0.05).

View Article: PubMed Central - PubMed

Affiliation: School of Veterinary Medicine, UCD Dublin, Ireland ; Food for Health Ireland (FHI), UCD Belfield, Dublin, Ireland.

ABSTRACT
Bioactive milk peptides are reported to illicit a range of physiological benefits and have been proposed as potential functional food ingredients. The objective of this study was to characterize the anti-inflammatory properties of sodium caseinate (NaCAS), its enzyme hydrolysate (EH) and peptide-enriched fractions (5 kDa retentate [R], 1 kDaR and 1 kDa permeate [P]), both in vitro using a Caco-2 cell line, and also ex vivo using a porcine colonic tissue explant system. Caco-2 cells were stimulated with tumour necrosis factor alpha (TNFα) and co-treated with casein hydrolysates for 24 h. Following this, interleukin (IL)-8 concentrations in the supernatant were measured using enzyme-linked immunosorbent assay. Porcine colonic tissue was stimulated with lipopolysaccharide and co-treated with casein hydrolysates for 3 h. The expression of a panel of inflammatory cytokines was measured using qPCR. While dexamethasone reduced the IL-8 concentration by 41.6%, the 1 kDaR and 1 kDaP fractions reduced IL-8 by 68.7% and 66.1%, respectively, relative to TNFα-stimulated Caco-2 cells (P < 0.05). In the ex vivo system, only the 1 kDaR fraction elicited a decrease inIL1-α,IL1-β,IL-8,TGF-β andIL-10 expression (P < 0.05). This study provides evidence that the bioactive peptides present in the 1 kDaR fraction of the NaCAS hydrolysate possess anti-inflammatory properties in vitro and ex vivo. Further in vivo analysis of the anti-inflammatory properties of the 1 kDaR is proposed.

No MeSH data available.


Related in: MedlinePlus