Human Milk Oligosaccharides Activate Epidermal Growth Factor Receptor and Protect Against Hypoxia-Induced Injuries in the Mouse Intestinal Epithelium and Caco2 Cells.

Abstract

BACKGROUND:

Hypoxia-induced intestinal barrier injuries lead to necrotizing enterocolitis (NEC). Although NEC in preterm neonates is preventable by human milk oligosaccharides (HMOs), the underlying mechanism remains unknown.

OBJECTIVE:

To reveal the role and mechanism of HMOs in protecting against hypoxia-induced injuries in intestinal epithelium of neonatal mice and cultured Caco2 cells.

METHODS:

NEC was induced by hypoxia and cold stress. Seventy C57BL/C pups (7-d-old) were divided into 5 groups and fed maternal breast milk (BM), formula alone (FF), or the formula added with HMOs at 5 (LHMO), 10 (MHMO), or 20 mg/mL (HHMO) for 3 d. Ileal hypoxia inducible factor 1α (HIF1α) and cleaved Caspase 3 were determined, along with staining for Ki-67 protein to labeled proliferative cells. In vitro, adherent Caco2 cells (undifferentiated, passage 14) were treated with HMOs, galacto-oligosaccharides, fructo-oligosaccharides, or mixed oligosaccharides at 10 mg/mL for 1 d exposed to 1% O2. Cell proliferation and apoptosis, along with phosphorylated epidermal growth factor receptor (P-EGFR) and 38KD MAPK (P-P38), were assayed in differentiated or undifferentiated Caco2 cells.

RESULTS:

Compared with the FF-fed mice, those fed MHMO and HHMO had 52% lower (P < 0.05) NEC scores, 60-80% greater (P < 0.05) KI67-positive cell numbers, and 56-71% decreases (P < 0.05) in ileal HIF1α and cleaved Caspase 3 (56-71%). Compared with those untreated, the HMO-treated Caco2 cells displayed 60% greater (P < 0.05) proliferative activity and 19% lower (P < 0.05) apoptotic cells after the hypoxia exposure. The HMO treatment led to 58% or 10-fold increases (P < 0.05) of P-EGFR and 48-89% decreases (P < 0.05) of P-P38 in either differentiated or undifferentiated Caco2 cells compared with the controls.

CONCLUSION:

Supplementing HMOs at 10-20 mg/mL into the formula for neonatal mice or media for Caco2 cells conferred protection against the hypoxia-induced injuries. The protection in the Caco2 cells was associated with an activation of EGFR.

• Authors: Chenyuan Wang, Ming Zhang, Huiyuan Guo, Jingyu Yan, Lingli Chen, Wendi Teng, Fazheng Ren, Yiran Li, Xifan Wang, Jie Luo, Yixuan Li
• Journal: The Journal of nutrition (J Nutr) Vol. 150 Issue 4 Pg. 756-762 (04 01 2020) ISSN: 1541-6100 [Electronic] United States
• PMID: 31915826 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)
• Copyright: Copyright © The Author(s) 2020.

Chemical References

• Oligosaccharides
• ErbB Receptors
• p38 Mitogen-Activated Protein Kinases

Topics

• Animals
• Animals, Newborn
• Apoptosis (drug effects)
• Caco-2 Cells
• Cell Proliferation (drug effects)
• Enterocolitis, Necrotizing (etiology, pathology, prevention & control)
• ErbB Receptors (drug effects, metabolism)
• Female
• Humans
• Hypoxia (complications, pathology)
• Intestinal Mucosa (drug effects, pathology)
• Mice
• Mice, Inbred C57BL
• Milk, Human (chemistry)
• Oligosaccharides (administration & dosage)
• Phosphorylation (drug effects)
• p38 Mitogen-Activated Protein Kinases (drug effects, metabolism)