Limits...
Alanyl-glutamine attenuates 5-fluorouracil-induced intestinal mucositis in apolipoprotein E-deficient mice.

Araújo CV, Lazzarotto CR, Aquino CC, Figueiredo IL, Costa TB, Alves LA, Ribeiro RA, Bertolini LR, Lima AA, Brito GA, Oriá RB - Braz. J. Med. Biol. Res. (2015)

Bottom Line: Apolipoprotein E (APOE=gene, apoE=protein) is a known factor regulating the inflammatory response that may have regenerative effects during tissue recovery from injury.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.Altogether our findings suggest APOE-independent Ala-Gln regenerative effects after 5-FU challenge.

View Article: PubMed Central - PubMed

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.

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.

Show MeSH

Related in: MedlinePlus

Mean mitotic (A) and apoptotic (B)indexes of H&E-stained ileal crypts from C57BL6J ApoE-knockout(APOE-/-) and wild-type (APOE+/+) mice following5-fluorouracil (5-FU)-induced intestinal mucositis (450 mg/kg,ip, single dose) and treated with the alanyl-glutamine(Ala-Gln, 100 mM) or phosphate buffered saline (PBS, control) via gavage. Atleast 10 crypts per animal were analyzed (n=4 animals/group) on the 6th daypost-challenge. Data are reported as means±SEM. aP<0.001vs unchallenged APOE+/+ mice;bP<0.05 vs unchallenged APOE-/- mice(one-way ANOVA and the Bonferroni test).
© Copyright Policy - open-access
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC4470307&req=5

f04: Mean mitotic (A) and apoptotic (B)indexes of H&E-stained ileal crypts from C57BL6J ApoE-knockout(APOE-/-) and wild-type (APOE+/+) mice following5-fluorouracil (5-FU)-induced intestinal mucositis (450 mg/kg,ip, single dose) and treated with the alanyl-glutamine(Ala-Gln, 100 mM) or phosphate buffered saline (PBS, control) via gavage. Atleast 10 crypts per animal were analyzed (n=4 animals/group) on the 6th daypost-challenge. Data are reported as means±SEM. aP<0.001vs unchallenged APOE+/+ mice;bP<0.05 vs unchallenged APOE-/- mice(one-way ANOVA and the Bonferroni test).

Mentions: Albeit we found enhanced mitotic crypt index 6 days-following 5-FU-injection, thiseffect was not significantly different compared to the unchallenged controls both inthe wild-type and APOE(-/-) mice. Ala-Gln was not able to improve cryptmitotic index compared with the untreated challenged mice in both mouse strains.However, Ala-Gln improved crypt mitotic index only when compared with theunchallenged control (p <0.01). 5-FU challenge remarkably increased crypt apopoticindex regardless of the mouse genetic background (P <0.05). Ala-Gln was unable toreduce the crypt apoptotic index following 5-FU challenge (Figure 4).


Alanyl-glutamine attenuates 5-fluorouracil-induced intestinal mucositis in apolipoprotein E-deficient mice.

Araújo CV, Lazzarotto CR, Aquino CC, Figueiredo IL, Costa TB, Alves LA, Ribeiro RA, Bertolini LR, Lima AA, Brito GA, Oriá RB - Braz. J. Med. Biol. Res. (2015)

Mean mitotic (A) and apoptotic (B)indexes of H&E-stained ileal crypts from C57BL6J ApoE-knockout(APOE-/-) and wild-type (APOE+/+) mice following5-fluorouracil (5-FU)-induced intestinal mucositis (450 mg/kg,ip, single dose) and treated with the alanyl-glutamine(Ala-Gln, 100 mM) or phosphate buffered saline (PBS, control) via gavage. Atleast 10 crypts per animal were analyzed (n=4 animals/group) on the 6th daypost-challenge. Data are reported as means±SEM. aP<0.001vs unchallenged APOE+/+ mice;bP<0.05 vs unchallenged APOE-/- mice(one-way ANOVA and the Bonferroni test).
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4470307&req=5

f04: Mean mitotic (A) and apoptotic (B)indexes of H&E-stained ileal crypts from C57BL6J ApoE-knockout(APOE-/-) and wild-type (APOE+/+) mice following5-fluorouracil (5-FU)-induced intestinal mucositis (450 mg/kg,ip, single dose) and treated with the alanyl-glutamine(Ala-Gln, 100 mM) or phosphate buffered saline (PBS, control) via gavage. Atleast 10 crypts per animal were analyzed (n=4 animals/group) on the 6th daypost-challenge. Data are reported as means±SEM. aP<0.001vs unchallenged APOE+/+ mice;bP<0.05 vs unchallenged APOE-/- mice(one-way ANOVA and the Bonferroni test).
Mentions: Albeit we found enhanced mitotic crypt index 6 days-following 5-FU-injection, thiseffect was not significantly different compared to the unchallenged controls both inthe wild-type and APOE(-/-) mice. Ala-Gln was not able to improve cryptmitotic index compared with the untreated challenged mice in both mouse strains.However, Ala-Gln improved crypt mitotic index only when compared with theunchallenged control (p <0.01). 5-FU challenge remarkably increased crypt apopoticindex regardless of the mouse genetic background (P <0.05). Ala-Gln was unable toreduce the crypt apoptotic index following 5-FU challenge (Figure 4).

Bottom Line: Apolipoprotein E (APOE=gene, apoE=protein) is a known factor regulating the inflammatory response that may have regenerative effects during tissue recovery from injury.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.Altogether our findings suggest APOE-independent Ala-Gln regenerative effects after 5-FU challenge.

View Article: PubMed Central - PubMed

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.

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.

Show MeSH
Related in: MedlinePlus