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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 and NaCAS enzyme hydrolysate, retentates and permeate on relative quantity (RQ) of genes un-stimulated by LPS challenge for 3 h in porcine colonic tissue explant. After 3 h of treatment with milk hydrolysates, total RNA was extracted, cDNA synthesised and RT-qPCR performed to quantify the selected panel of genes. RQ values of treatments with NaCAS, EH, 5 kDaR, 1 kDaR or 1 kDaP compared to unchallenged control, error bars indicate SE. *P < 0.05, **P < 0.01.
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fig04: Effect of NaCAS and NaCAS enzyme hydrolysate, retentates and permeate on relative quantity (RQ) of genes un-stimulated by LPS challenge for 3 h in porcine colonic tissue explant. After 3 h of treatment with milk hydrolysates, total RNA was extracted, cDNA synthesised and RT-qPCR performed to quantify the selected panel of genes. RQ values of treatments with NaCAS, EH, 5 kDaR, 1 kDaR or 1 kDaP compared to unchallenged control, error bars indicate SE. *P < 0.05, **P < 0.01.

Mentions: While the LPS challenge had no effect on the expression ofIL-10,IL-17 andTGF-β, as presented in Figure2, co-treatment with casein hydrolysates had significant effects onIL-10 andTGF-β expression, whereas co-treatment with NaCAS had significant effects on onlyIL-17 expression relative to unchallenged tissues, presented in Figure4. The treatment of porcine colonic tissues with EH, 5 kDaR, 1 kDaR and 1 kDaP caused a down-regulation ofIL-10 expression (0.083, 0.056, 0.063, 0.098 vs. 0.194 ± 0.029,P < 0.05) relative to unchallenged tissues. Colonic tissues co-treated with NaCAS was associated with an increase inIL-17 (0.358 vs. 0.136 ± 0.042,P < 0.05) expression relative to unchallenged tissues. The co-treatment with EH, 5 kDaR and 1 kDaR hydrolysates was associated with a down-regulation ofTGF-β (0.121, 0.181, 0.105 vs. 0.382 ± 0.071,P < 0.05) expression in colonic tissues relative to unchallenged 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 and NaCAS enzyme hydrolysate, retentates and permeate on relative quantity (RQ) of genes un-stimulated by LPS challenge for 3 h in porcine colonic tissue explant. After 3 h of treatment with milk hydrolysates, total RNA was extracted, cDNA synthesised and RT-qPCR performed to quantify the selected panel of genes. RQ values of treatments with NaCAS, EH, 5 kDaR, 1 kDaR or 1 kDaP compared to unchallenged 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

fig04: Effect of NaCAS and NaCAS enzyme hydrolysate, retentates and permeate on relative quantity (RQ) of genes un-stimulated by LPS challenge for 3 h in porcine colonic tissue explant. After 3 h of treatment with milk hydrolysates, total RNA was extracted, cDNA synthesised and RT-qPCR performed to quantify the selected panel of genes. RQ values of treatments with NaCAS, EH, 5 kDaR, 1 kDaR or 1 kDaP compared to unchallenged control, error bars indicate SE. *P < 0.05, **P < 0.01.
Mentions: While the LPS challenge had no effect on the expression ofIL-10,IL-17 andTGF-β, as presented in Figure2, co-treatment with casein hydrolysates had significant effects onIL-10 andTGF-β expression, whereas co-treatment with NaCAS had significant effects on onlyIL-17 expression relative to unchallenged tissues, presented in Figure4. The treatment of porcine colonic tissues with EH, 5 kDaR, 1 kDaR and 1 kDaP caused a down-regulation ofIL-10 expression (0.083, 0.056, 0.063, 0.098 vs. 0.194 ± 0.029,P < 0.05) relative to unchallenged tissues. Colonic tissues co-treated with NaCAS was associated with an increase inIL-17 (0.358 vs. 0.136 ± 0.042,P < 0.05) expression relative to unchallenged tissues. The co-treatment with EH, 5 kDaR and 1 kDaR hydrolysates was associated with a down-regulation ofTGF-β (0.121, 0.181, 0.105 vs. 0.382 ± 0.071,P < 0.05) expression in colonic tissues relative to unchallenged 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