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Title in Portuguese: Glutamine and Alanyl-Glutamine increase RhoA expression and reduce clostridium difficile Toxin-A-Induced intestinal epithelial cell damage
Author: Santos, Ana Angélica Queiroz Assunção
Braga Neto, Manuel Bonfim
Oliveira, Marcelo Róseo de
Freire, Rosemayre Souza
Barros, Eduardo Bedê
Santiago, Thiago de Melo
Alencar, Luciana Magalhães Rebêlo
Mermelstein, Claudia
Warren, Cirle A.
Guerrant, Richard L.
Brito, Gerly Anne de Castro
Keywords: Microscopy
Issue Date: 2013
Publisher: BioMed Research International
Citation: SANTOS, Ana Angélica Queiroz Assunção; BRAGA-NETO, Manuel B; OLIVEIRA, Marcelo Róseo de; FREIRE, Rosemayre Souza; BARROS, Eduardo Bedê; SANTIAGO, Thiago de Melo; ALENCAR, Luciana Magalhães Rebêlo; MERMELSTEIN, Claudia; WARREN, Cirle A; GUERRANT, Richard L; BRITO, Gerly Anne de Castro. Glutamine and Alanyl-Glutamine increase RhoA expression and reduce clostridium difficile Toxin-A-Induced intestinal epithelial cell damage. BioMed Research International, v. 2013, p. 1-14, 2013.
Abstract: Clostridium difficile is a major cause of antibiotic-associated colitis and is associated with signi􀄕cant morbidity and mortality. Glutamine (Gln) is a major fuel for the intestinal cell population. Alanyl-glutamine (Ala-Gln) is a dipeptide that is highly soluble and well tolerated. IEC-6 cells were used in the in vitro experiments. Cell morphology was evaluated by atomic force microscopy (AFM) and scanning electron microscopy (SEM). Cell proliferation was assessed by WST-1 and Ki-67 and apoptosis was assessed by TUNEL. Cytoskeleton was evaluated by immuno􀄘uorescence for RhoA and F-actin. RhoA was quanti􀄕ed by immunoblotting. TcdA induced cell shrinkage as observed by AFM, SEM, and 􀄘uorescent microscopy. Additionally, collapse of the F-actin cytoskeleton was demonstrated by immuno􀄘uorescence. TcdA decreased cell volume and area and increased cell height by 79%, 66.2%, and 58.9%, respectively. Following TcdA treatment, Ala-Gln and Gln supplementation, signi􀄕cantly increased RhoA by 65.5% and 89.7%, respectively at 24 h. Ala-Gln supplementation increased cell proliferation by 137.5% at 24 h and decreased cell apoptosis by 61.4% at 24 h following TcdA treatment. In conclusion, TcdA altered intestinal cell morphology and cytoskeleton organization, decreased cell proliferation, and increased cell apoptosis. Ala-Gln and Gln supplementation reduced intestinal epithelial cell damage and increased RhoA expression.
metadata.dc.type: Artigo de Periódico
ISSN: 2314-6133
Appears in Collections:DFI - Artigos publicados em revista científica

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