title: Bovine lactoferrin regulates cell survival, apoptosis and inflammation in intestinal epithelial cells and preterm pig intestine.
abstract: Bovine lactoferrin (bLF) may modulate neonatal intestinal inflammation. Previous studies in intestinal epithelial cells (IECs) indicated that moderate bLF doses enhance proliferation whereas high doses trigger inflammation. To further elucidate cellular mechanisms, we profiled the porcine IEC proteome after stimulation with bLF at 0, 0.1, 1 and 10g/L by LC-MS-based proteomics. Key pathways were analyzed in the intestine of formula-fed preterm pigs with and without supplementation of 10g/L bLF. Levels of 123 IEC proteins were altered by bLF. Low bLF doses (0.1-1g/L) up-regulated 11 proteins associated with glycolysis, energy metabolism and protein synthesis, indicating support of cell survival. In contrast, a high bLF dose (10g/L) up-regulated three apoptosis-inducing proteins, down-regulated five anti-apoptotic and proliferation-inducing proteins and 15 proteins related to energy and amino acid metabolism, and altered three proteins enhancing the hypoxia inducible factor-1 (HIF-1) pathway. In the preterm pig intestine, bLF at 10g/L decreased villus height/crypt depth ratio and up-regulated the Bax/Bcl-2 ratio and HIF-1α, indicating elevated intestinal apoptosis and inflammation. In conclusion, bLF dose-dependently affects IECs via metabolic, apoptotic and inflammatory pathways. It is important to select an appropriate dose when feeding neonates with bLF to avoid detrimental effects exerted by excessive doses.
  
  
  CONCLUSIONS
  The present work elucidates dose-dependent effects of bLF on the proteomic changes of IECs in vitro supplemented with data from a preterm pig study confirming detrimental effects of enteral feeding with the highest dose of bLF (10g/L). The study contributes to further understanding on mechanisms that bLF, as an important milk protein, can regulate the homeostasis of the immature intestine. Results from this study urge neonatologists to carefully consider the dose of bLF to supplement into infant formula used for preterm neonates.
authors.affiliation: Comparative Pediatrics and Nutrition, Department of Veterinary Clinical and Animal Sciences, University of Copenhagen, DK-1958, Denmark; Department of Food Science, University of Copenhagen, DK-1958, Denmark.
authors.forename: Duc Ninh
authors.initials: DN
authors.lastname: Nguyen
authors.affiliation: Comparative Pediatrics and Nutrition, Department of Veterinary Clinical and Animal Sciences, University of Copenhagen, DK-1958, Denmark.
authors.forename: Pingping
authors.initials: P
authors.lastname: Jiang
authors.affiliation: Department of Health Science and Technology, Aalborg University, DK-9220, Denmark.
authors.forename: Allan
authors.initials: A
authors.lastname: Stensballe
authors.affiliation: Department of Molecular Biology and Genetics, Aarhus University, DK-8000, Denmark.
authors.forename: Emøke
authors.initials: E
authors.lastname: Bendixen
authors.affiliation: Comparative Pediatrics and Nutrition, Department of Veterinary Clinical and Animal Sciences, University of Copenhagen, DK-1958, Denmark.
authors.forename: Per T
authors.initials: PT
authors.lastname: Sangild
authors.affiliation: Comparative Pediatrics and Nutrition, Department of Veterinary Clinical and Animal Sciences, University of Copenhagen, DK-1958, Denmark; Department of Food Science, University of Copenhagen, DK-1958, Denmark. Electronic address: dcha@food.ku.dk.
authors.forename: Dereck E W
authors.initials: DE
authors.lastname: Chatterton
mesh_terms: D000818:Animals; D017209:Apoptosis / Q000187:drug effects*; D051017:Apoptosis Regulatory Proteins / Q000378:metabolism*; D002417:Cattle; D018797:Cell Cycle Proteins / Q000378:metabolism*; D002470:Cell Survival / Q000187:drug effects; D004847:Epithelial Cells / Q000378:metabolism* / Q000473:pathology; D007249:Inflammation / Q000378:metabolism / Q000473:pathology; D007413:Intestinal Mucosa / Q000378:metabolism* / Q000473:pathology; D007781:Lactoferrin / Q000494:pharmacology*; D013552:Swine
chemical_list: D051017:Apoptosis Regulatory Proteins; D018797:Cell Cycle Proteins; D007781:Lactoferrin
keywords: Apoptosis; Inflammation; Lactoferrin; Necrotizing enterocolitis