Human Milk Exosomes Induce ZO-1 Expression via Inhibition of REDD1 Expression in Human Intestinal Epithelial Cells

Human milk exosomes (HMEs) enhance intestinal barrier function and contribute to an improvement in inflammation and mucosal injury, such as necrotizing enteritis (NEC), in infants. Here, we aimed to elucidate the intracellular factors involved in HME-induced expression of zonula occludens-1 (ZO-1), a tight junction protein, in Caco-2 human intestinal epithelial cells. HME treatment for 72 h significantly increased transepithelial electrical resistance in these cells. The mean ZO-1 protein levels in cells treated with HME for 72 h were significantly higher than those in the control cells. The mRNA and protein levels of regulated in development and DNA damage response 1 (REDD1) in HME-treated cells were significantly lower than those in the control cells. Although HME treatment did not increase the mechanistic target of rapamycin (mTOR) level in Caco-2 cells, it significantly increased the phosphorylated mTOR (p-mTOR) level and p-mTOR/mTOR ratio. The ZO-1 protein levels in cells treated with an inducer of REDD1, cobalt chloride (CoCl₂) alone were significantly lower than those in the control cells. However, ZO-1 protein levels in cells co-treated with HME and CoCl₂ were significantly higher than those in cells treated with CoCl₂ alone. Additionally, REDD1 protein levels in cells treated with CoCl₂ alone were significantly higher than those in the control cells. However, REDD1 protein levels in cells co-treated with HME and CoCl₂ were significantly lower than those in cells treated with CoCl₂ alone. This HME-mediated effect may contribute to the development of barrier function in the infant intestine and protect infants from diseases.