Journal
JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY
Volume 67, Issue 19, Pages 5634-5646Publisher
AMER CHEMICAL SOC
DOI: 10.1021/acs.jafc.9b01324
Keywords
infant formula; protein oxidation; advanced-glycation endproducts; D-amino acids; Caco-2 cells
Funding
- Novo Nordisk Foundation [NNF130C0004294]
- Irish Department of Agriculture and Food (FIRM grant) [15-F-604 TOMI]
- Chinese Scholarship Council (CSC) [201506170056]
- Novo Nordisk Fonden [NNF13OC0004294] Funding Source: researchfish
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Proteins present in infant formulas are modified by oxidation and glycation during processing. Modified amino acid residues released from proteins may be absorbed in the gastrointestinal tract, and pose a health risk to infants. In this study, the markers of glycation furosine (1.7-3.5, mu g per milligram of protein) and N-epsilon-(carboxymethy)lysine (28-81 ng per milligram of protein) were quantitated in infant formulas. The effects of these species, and other amino acid modifications, at the levels detected in infant formulas, on 3T3-L1 (murine preadipocyte) and Caco-2 (human intestinal epithelial) cells were assessed. Incubation of 3T3-L1 cells for 48 h with amino acid side chain oxidation and glycation products (1 and 10 mu M) resulted in a loss (up to 40%, p < 0.05) of cell thiols and decreased metabolic activity compared with those of the controls. In contrast, Caco-2 cells showed a stimulation (10-50%, p < 0.05) of cellular metabolism on exposure to these products for 24 or 48 h. A 28% (p < 0.05) increase in protein carbonyls was detected upon incubation with 200 mu M modified amino acids for 48 h, although no alteration in transepithelial electrical resistance was detected. Oxidation products were detected in the basolateral compartments of Caco-2 monolayers when modified amino acids were applied to the apical side, consistent with limited permeability (up to 3.4%) across the monolayer. These data indicate that modified amino acids present in infant formulas can induce effects on different cell types, with evidence of bioavailability and induction of cellular stress. This may lead to potential health risks for infants consistently exposed to high levels of infant formulas.
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