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Glutamine and Alanyl-Glutamine increase RhoA expression and reduce clostridium difficile Toxin-A-Induced intestinal epithelial cell damage
- Source :
- Repositório Institucional da Universidade Federal do Ceará (UFC), Universidade Federal do Ceará (UFC), instacron:UFC
- Publication Year :
- 2013
- Publisher :
- BioMed Research International, 2013.
-
Abstract
- Clostridium difficile is a major cause of antibiotic-associated colitis and is associated with signicant 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 immunouorescence for RhoA and F-actin. RhoA was quantied by immunoblotting. TcdA induced cell shrinkage as observed by AFM, SEM, and uorescent microscopy. Additionally, collapse of the F-actin cytoskeleton was demonstrated by immunouorescence. 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, signicantly 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.
- Subjects :
- Microscopy
Cell
Cytoskeleton
Subjects
Details
- Language :
- English
- Database :
- OpenAIRE
- Journal :
- Repositório Institucional da Universidade Federal do Ceará (UFC), Universidade Federal do Ceará (UFC), instacron:UFC
- Accession number :
- edsair.od......3056..ba40dc01a08e4534e12604af0046776e