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

Chenyuan Wang; Ming Zhang; Huiyuan Guo; Jingyu Yan; Lingli Chen; Wendi Teng; Fazheng Ren; Yiran Li; Xifan Wang; Jie Luo; Yixuan Li

doi:10.1093/jn/nxz297

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

J Nutr. 2020 Apr 1;150(4):756-762. doi: 10.1093/jn/nxz297.

Authors

Chenyuan Wang^{1

2}, Ming Zhang³, Huiyuan Guo^{1

2}, Jingyu Yan⁴, Lingli Chen^{1

2}, Wendi Teng^{1

2}, Fazheng Ren^{1

2}, Yiran Li^{1

2}, Xifan Wang^{1

2}, Jie Luo⁵, Yixuan Li¹

Affiliations

¹ Beijing Advanced Innovation Center for Food Nutrition and Human Health, College of Food Science & Nutritional Engineering, China Agricultural University, Beijing, China.
² Key Laboratory of Functional Dairy, China Agricultural University, Beijing, China.
³ School of Food and Chemical Engineering, Beijing Technology and Business University, Beijing, China.
⁴ Dalian Institute of Chemical Physics, Chinese Academy of Sciences, Dalian, China.
⁵ College of Food Science and Technology, Hunan Agricultural University, Changsha, China.

PMID: 31915826
DOI: 10.1093/jn/nxz297

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.

Keywords: EGFR; P38; human milk oligosaccharides; hypoxia; necrotizing enterocolitis.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

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

Substances

Oligosaccharides
ErbB Receptors
p38 Mitogen-Activated Protein Kinases