title: Apolipoprotein E COG 133 mimetic peptide improves 5-fluorouracil-induced intestinal mucositis.
abstract: BACKGROUND
  Intestinal mucositis is one of the major troublesome side effects of anticancer chemotherapy leading to poor patient compliance. In this study we addressed the role of the novel apolipoprotein E (ApoE) COG 133 mimetic peptide in 5-fluorouracil (5-FU)-challenged Swiss mice and IEC-6 cell monolayers. Experiments were also conducted in C57BL6J ApoE knock-out mice to assess the effects of apoE peptide treatment.
  
  
  METHODS
  Experimental groups were as follows: unchallenged controls, 5-FU-challenged mice (450 mg/kg, i.p) with or without the ApoE peptide (0.3, 1, and 3 μM, given twice daily i.p. for 4 days). Mice were sacrificed 3 days after 5-FU challenge. Proximal small intestinal samples were harvested for molecular biology and histological processing. We conducted ELISA assays and RT-PCR to target IL-1β, TNF-α, IL-10, iNOS, and myeloperoxidase (MPO) to assess intestinal inflammation. Cell death and NF-κB assays were also conducted in apoE knock-out mice. In our in vitro models, IEC-6 cells were exposed to 1 mM of 5-FU in glutamine free media with or without the ApoE peptide (0.02, 0.2, 2, 5, 10, and 20 μM). We investigated IEC-6 cell proliferation and migration, 24 h after the 5-FU challenge. Additionally, apoptotic IEC-6 cells were measured by Tunel and flow cytometry. Equimolar doses of the ApoA-I (D4-F) peptide were also used in some experiments for comparative studies.
  
  
  RESULTS
  Villus blunting and heavy inflammatory infiltrates were seen in the 5-FU-challenged group, findings that were partially ameliorated by the ApoE peptide. We found increased intestinal MPO and pro-inflammatory IL-1β and TNF-α levels, and TNF-α and iNOS transcripts, and reduction of IL-10 following 5-FU treatment, each of which were partially abrogated by the peptide. Improvements were also found in IEC-6 cell apoptosis and migration following ApoE and D-4F treatment.
  
  
  CONCLUSIONS
  Altogether, these findings suggest that the novel ApoE COG 133 mimetic peptide can reduce 5-FU-induced intestinal changes and potentially benefit mucositis.
authors.affiliation: Center for Global Health, School of Medicine, University of Virginia, Carter Harrison Bldg MR-6, 625 Crispell Drive, Charlottesville, VA 22908, USA.
authors.forename: Orleâncio Gomes R
authors.initials: OG
authors.lastname: Azevedo
authors.forename: Renato André C
authors.initials: RA
authors.lastname: Oliveira
authors.forename: Bruna Castro
authors.initials: BC
authors.lastname: Oliveira
authors.forename: Snjezana
authors.initials: S
authors.lastname: Zaja-Milatovic
authors.forename: Celina Viana
authors.initials: CV
authors.lastname: Araújo
authors.forename: Deysi Viviana T
authors.initials: DV
authors.lastname: Wong
authors.forename: Tiê Bezerra
authors.initials: TB
authors.lastname: Costa
authors.forename: Herene Barros Miranda
authors.initials: HB
authors.lastname: Lucena
authors.forename: Roberto César P
authors.initials: RC
authors.lastname: Lima
authors.forename: Ronaldo A
authors.initials: RA
authors.lastname: Ribeiro
authors.forename: Cirle A
authors.initials: CA
authors.lastname: Warren
authors.forename: Aldo Ângelo M
authors.initials: AÂ
authors.lastname: Lima
authors.forename: Michael P
authors.initials: MP
authors.lastname: Vitek
authors.forename: Richard L
authors.initials: RL
authors.lastname: Guerrant
authors.forename: Reinaldo B
authors.initials: RB
authors.lastname: Oriá
mesh_terms: D000818:Animals; D001057:Apolipoproteins E / Q000235:genetics / Q000494:pharmacology / Q000627:therapeutic use*; D017209:Apoptosis / Q000187:drug effects; D040761:Biomimetic Materials / Q000494:pharmacology / Q000627:therapeutic use; D002465:Cell Movement / Q000187:drug effects; D049109:Cell Proliferation / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D016207:Cytokines / Q000378:metabolism; D004195:Disease Models, Animal; D005472:Fluorouracil / Q000009:adverse effects*; D066298:In Vitro Techniques; D007410:Intestinal Diseases / Q000139:chemically induced* / Q000188:drug therapy* / Q000473:pathology; D007583:Jejunum / Q000187:drug effects / Q000378:metabolism / Q000473:pathology; D051379:Mice; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D052016:Mucositis / Q000139:chemically induced* / Q000188:drug therapy* / Q000473:pathology; D010446:Peptide Fragments / Q000494:pharmacology / Q000627:therapeutic use; D009195:Peroxidase / Q000378:metabolism; D051381:Rats
chemical_list: D001057:Apolipoproteins E; D016207:Cytokines; D010446:Peptide Fragments; C579624:apolipoprotein E (133-149); D009195:Peroxidase; D005472:Fluorouracil