Protective Effects of Human Milk-Derived Exosomes on Intestinal Stem Cells Damaged by Oxidative Stress

Abstract

Breastfeeding has been shown to have a protective effect on the occurrence of necrotizing enterocolitis (NEC), but the mechanism remains unclear. In the context of NEC pathogenesis, many of the protective properties of exosomes on the intestinal epithelial compartment make it an ideal therapeutic target. In the present study, our hypothesis was that intestinal stem cells (ISCs) would be protected from injury by human milk-derived exosomes (HMDEs). Human breast milk was collected, and exosomes were isolated using ExoQuick reagent. Magnetic-activated cell sorting isolation of prominin-1+ ISCs was performed from small intestines of neonatal rat. ISCs were treated with or without H2O2, and HMDEs, an equal volume of HMDE-free milk, or a control solution [phosphate-buffered solution (PBS)] was added, respectively. In the absence of HMDEs, exposure of ISCs to H2O2 led to decreased cell viability. However, addition of HMDEs to ISCs exposed to H2O2 led to significantly increased ISC viability. There was a significant upregulation of mRNA expression of Axin2, c-Myc, and Cyclin D1 genes of the Wnt/β-catenin axis in ISCs treated with HMDEs (6.99 ± 2.34, 4.21 ± 1.68, 6.17 ± 2.22, respectively, P < 0.05 for all), as compared to control. In the presence of carnosic acid (a specific Wnt/β-catenin signaling inhibitor), the cell viability was significantly decreased. Thus, HMDEs protect ISCs from oxidative stress injury in vitro, which were possibly mediated via the Wnt/β-catenin signaling pathway. Our findings indicate that oral administration of HMDEs might be a promising measure in treating NEC or in preventing the development of NEC in high-risk infants when breast milk is not available.

Porcine Milk-Derived Small Extracellular Vesicles Promote Intestinal Immunoglobulin Production through pIgR

“…Many studies have confirmed that milk sEVs have a variety of biological functions, including intestinal health and immune regulations. Milk sEVs promote the growth of IECs [ 15 ] and reduce intestinal injury induced by oxidative stress [ 16 , 17 ], intestinal inflammation [ 33 ], and necrotizing enterocolitis [ 18 , 34 ]. A recent publication showed that camel milk-derived sEVs ameliorate cyclophosphamide-induced immunotoxicity in rats [ 35 ].…”

“…Due to the protective action of the lipid bilayer of milk sEVs, their ncRNA cargos are stable and resistant to harsh conditions, including gastrointestinal digestive juices, freeze–thaw cycles, and RNase digestion [ 12 , 13 , 14 ]. Studies have shown that milk sEVs promote the growth of IECs [ 15 ] and reduce intestinal injury induced by oxidative stress [ 16 , 17 ] and necrotizing enterocolitis [ 18 ]. Milk sEVs-derived RNAs can be absorbed by macrophages and intestinal cells in vitro [ 19 , 20 ], and can enter the circulatory system of milk consumers in vivo [ 21 ].…”

Maternal Microbiota, Early Life Colonization and Breast Milk Drive Immune Development in the Newborn

“…The treatment with milk-derived exosomes prevented intestinal injury and the reduction of goblet cells, which is a hallmark of NEC ( 236 ). Lately, human milk-derived exosomes attenuated intestinal damage and protected intestinal stem cells from undergoing apoptosis due to oxidative stress in vitro ( 237 , 238 ). Altogether, breast milk-derived exosomes seem to protect the mucosal environment against injury and inflammation-mediated cell death by positively affecting different cell types of the intestinal epithelium.…”

Stem cells and exosomes: promising candidates for necrotizing enterocolitis therapy

“… / [ 49 ] HM-Ex Human ExoQuick reagent 0.5 mg/mL / In vitro / HM-Exs upregulate Wnt/β-catenin signaling in ISCs and increase cell viability under H 2 O 2 exposure compared to the control group. Wnt/β-catenin (ISCs) [ 50 ] PM-Ex Pig Ultra-centrifugation 0.037 mg/μL Gavage In vitro In vivo Mouse pups with LPS (7.5 mg/kg, qd for 7 days) PM-Exs inhibit intestinal epithelial cell apoptosis and decrease TLR4/NF-κB signaling through miRNAs in vitro. PM-Exs prevent LPS-induced intestinal injury and inflammation in vivo.…”

“…result in further reduction of NEC incidences compared to those administered intraperitoneally, which indicates that the enteral route is a better option for HM-Ex administration [ 48 ]. The mechanisms by which HM-Exs reduce NEC incidences are partially contributed by activation of the Wnt/β-catenin signaling pathway, which increases ISC viability and protects cells from oxidative stress [ 50 ].…”