Effects of human milk and spermine on hydrogen peroxide-induced oxidative damage in IEC-6 cells

Objective: Oxidative stress is intimately involved in the, pathologic processes of serious diseases in the perinatal period. Human milk (HM) contains various bioactive substances, some of which are known as antioxidants, including polyamines such as spermine (SPM). We examined the antioxidative properties of HM and SPM in an intestinal epithelial cell line. Method: Confluent Intestinal Epithelial Cells-6 (IEC-6) cells were preincubated with 100-fold dilutions of defatted HM, bovine milk, or three artificial milks for 24 hours, followed by hydrogen peroxide (H2O2) challenge (0.5 mM, 30 min) for oxidative stress. Cells were preincubated with either HM or increasing concentrations (within the range of HM) of SPM for 24 hours followed by an H2O2 challenge (0.25 mM, 30 min). Results: HM-treated cells showed the highest survival rate (50%) compared with no pretreatment (27%), bovine milk-treated (6%), or artificial formula-treated (13-16%) cells. Significantly higher survival rates were observed in the cells treated with HM (44.0%) and in those treated with 0.5, 1, or 5 mu M of SPM (12.6, 13.1, or 22.2%, respectively) in comparison with the nontreated cells (7.0%). Conclusion: Our results demonstrated that HM and SPM alleviated H2O2-induced oxidative damage in IEC-6 cells, whereas bovine milk and artificial formula did not show any anti-oxidative capacity. These results suggest that HM acts as an antioxidant in the gastrointestinal tract of infants and that SPM plays an important role in the antioxidative properties of HM.