title: Alanyl-glutamine attenuates 5-fluorouracil-induced intestinal mucositis in apolipoprotein E-deficient mice.
abstract: Apolipoprotein E (APOE=gene, apoE=protein) is a known factor regulating the inflammatory response that may have regenerative effects during tissue recovery from injury. We investigated whether apoE deficiency reduces the healing effect of alanyl-glutamine (Ala-Gln) treatment, a recognized gut-trophic nutrient, during tissue recovery after 5-FU-induced intestinal mucositis. APOE-knockout (APOE-/-) and wild-type (APOE+/+) C57BL6J male and female mice (N=86) were given either Ala-Gln (100 mM) or phosphate buffered saline (PBS) by gavage 3 days before and 5 days after a 5-fluorouracil (5-FU) challenge (450 mg/kg, via intraperitoneal injection). Mouse body weight was monitored daily. The 5-FU cytotoxic effect was evaluated by leukometry. Intestinal villus height, villus/crypt ratio, and villin expression were monitored to assess recovery of the intestinal absorptive surface area. Crypt length, mitotic, apoptotic, and necrotic crypt indexes, and quantitative real-time PCR for insulin-like growth factor-1 (IGF-1) and B-cell lymphoma 2 (Bcl-2) intestinal mRNA transcripts were used to evaluate intestinal epithelial cell turnover. 5-FU challenge caused significant weight loss and leukopenia (P<0.001) in both mouse strains, which was not improved by Ala-Gln. Villus blunting, crypt hyperplasia, and reduced villus/crypt ratio (P<0.05) were found in all 5-FU-challenged mice but not in PBS controls. Ala-Gln improved villus/crypt ratio, crypt length and mitotic index in all challenged mice, compared with PBS controls. Ala-Gln improved villus height only in APOE-/- mice. Crypt cell apoptosis and necrotic scores were increased in all mice challenged by 5-FU, compared with untreated controls. Those scores were significantly lower in Ala-Gln-treated APOE+/+ mice than in controls. Bcl-2 and IGF-1 mRNA transcripts were reduced only in the APOE-/- -challenged mice. Altogether our findings suggest APOE-independent Ala-Gln regenerative effects after 5-FU challenge.
authors.affiliation: Laboratório da Biologia da Cicatrização, Ontogenia e Nutrição de Tecidos, INCT - Instituto de Biomedicina do Semiárido Brasileiro, Faculdade de Medicina, Universidade Federal do Ceará, Fortaleza, CE, Brasil.
authors.forename: C V
authors.initials: CV
authors.lastname: Araújo
authors.affiliation: Laboratório de Biologia Molecular e do Desenvolvimento, Universidade de Fortaleza, Fortaleza, CE, Brasil.
authors.forename: C R
authors.initials: CR
authors.lastname: Lazzarotto
authors.affiliation: Laboratório da Biologia da Cicatrização, Ontogenia e Nutrição de Tecidos, INCT - Instituto de Biomedicina do Semiárido Brasileiro, Faculdade de Medicina, Universidade Federal do Ceará, Fortaleza, CE, Brasil.
authors.forename: C C
authors.initials: CC
authors.lastname: Aquino
authors.affiliation: Laboratório da Biologia da Cicatrização, Ontogenia e Nutrição de Tecidos, INCT - Instituto de Biomedicina do Semiárido Brasileiro, Faculdade de Medicina, Universidade Federal do Ceará, Fortaleza, CE, Brasil.
authors.forename: I L
authors.initials: IL
authors.lastname: Figueiredo
authors.affiliation: Laboratório da Biologia da Cicatrização, Ontogenia e Nutrição de Tecidos, INCT - Instituto de Biomedicina do Semiárido Brasileiro, Faculdade de Medicina, Universidade Federal do Ceará, Fortaleza, CE, Brasil.
authors.forename: T B
authors.initials: TB
authors.lastname: Costa
authors.affiliation: Laboratório da Biologia da Cicatrização, Ontogenia e Nutrição de Tecidos, INCT - Instituto de Biomedicina do Semiárido Brasileiro, Faculdade de Medicina, Universidade Federal do Ceará, Fortaleza, CE, Brasil.
authors.forename: L A de Oliveira
authors.initials: LA
authors.lastname: Alves
authors.affiliation: Laboratório da Inflamação e Câncer, INCT - Instituto de Biomedicina do Semiárido Brasileiro, Faculdade de Medicina, Universidade Federal do Ceará, Fortaleza, CE, Brasil.
authors.forename: R A
authors.initials: RA
authors.lastname: Ribeiro
authors.affiliation: Laboratório de Biologia Molecular e do Desenvolvimento, Universidade de Fortaleza, Fortaleza, CE, Brasil.
authors.forename: L R
authors.initials: LR
authors.lastname: Bertolini
authors.affiliation: Laboratório de Doenças Infecciosas, INCT - Instituto de Biomedicina do Semiárido Brasileiro, Faculdade de Medicina, Universidade Federal do Ceará, Fortaleza, CE, Brasil.
authors.forename: A A M
authors.initials: AA
authors.lastname: Lima
authors.affiliation: Laboratório da Inflamação e Câncer, INCT - Instituto de Biomedicina do Semiárido Brasileiro, Faculdade de Medicina, Universidade Federal do Ceará, Fortaleza, CE, Brasil.
authors.forename: G A C
authors.initials: GA
authors.lastname: Brito
authors.affiliation: Laboratório da Biologia da Cicatrização, Ontogenia e Nutrição de Tecidos, INCT - Instituto de Biomedicina do Semiárido Brasileiro, Faculdade de Medicina, Universidade Federal do Ceará, Fortaleza, CE, Brasil.
authors.forename: R B
authors.initials: RB
authors.lastname: Oriá
mesh_terms: D000818:Animals; D000964:Antimetabolites, Antineoplastic / Q000009:adverse effects*; D001057:Apolipoproteins E / Q000172:deficiency*; D017209:Apoptosis / Q000187:drug effects; D001835:Body Weight; D004151:Dipeptides / Q000494:pharmacology* / Q000627:therapeutic use; D005260:Female; D005472:Fluorouracil / Q000009:adverse effects*; D007334:Insulin-Like Growth Factor I / Q000032:analysis; D007413:Intestinal Mucosa / Q000187:drug effects* / Q000473:pathology; D007958:Leukocyte Count; D016393:Lymphoma, B-Cell; D008297:Male; D008810:Mice, Inbred C57BL; D008938:Mitosis / Q000187:drug effects; D052016:Mucositis / Q000139:chemically induced / Q000188:drug therapy* / Q000473:pathology; D011897:Random Allocation; D060888:Real-Time Polymerase Chain Reaction; D015203:Reproducibility of Results; D013997:Time Factors; D016896:Treatment Outcome
chemical_list: D000964:Antimetabolites, Antineoplastic; D001057:Apolipoproteins E; D004151:Dipeptides; D007334:Insulin-Like Growth Factor I; D005472:Fluorouracil; C054122:alanylglutamine