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Open Access Research article

Goat milk with and without increased concentrations of lysozyme improves repair of intestinal cell damage induced by enteroaggregative Escherichia coli

Eunice B Carvalho1, Elizabeth A Maga2, Josiane S Quetz1, Ila FN Lima1, Hemerson YF Magalhães1, Felipe AR Rodrigues1, Antônio VA Silva1, Mara MG Prata1, Paloma A Cavalcante1, Alexandre Havt1, Marcelo Bertolini2, Luciana R Bertolini2 and Aldo AM Lima13*

Author affiliations

1 Department of Physiology and Pharmacology & INCT-Biomedicine, Faculty of Medicine, Federal University of Ceará, Fortaleza, Ceará, Brazil

2 Department of Animal Science, University of California, Davis, CA, USA

3 School of Medicine, Health Science Center, University of Fortaleza, Fortaleza, Ceará, Brazil

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Citation and License

BMC Gastroenterology 2012, 12:106  doi:10.1186/1471-230X-12-106

Published: 11 August 2012

Abstract

Background

Enteroaggregative Escherichia coli (EAEC) causes diarrhea, malnutrition and poor growth in children. Human breast milk decreases disease-causing bacteria by supplying nutrients and antimicrobial factors such as lysozyme. Goat milk with and without human lysozyme (HLZ) may improve the repair of intestinal barrier function damage induced by EAEC. This work investigates the effect of the milks on intestinal barrier function repair, bacterial adherence in Caco-2 and HEp-2 cells, intestinal cell proliferation, migration, viability and apoptosis in IEC-6 cells in the absence or presence of EAEC.

Methods

Rat intestinal epithelial cells (IEC-6, ATCC, Rockville, MD) were used for proliferation, migration and viability assays and human colon adenocarcinoma (Caco-2, ATCC, Rockville, MD) and human larynx carcinoma (HEp-2, ATCC, Rockville, MD) cells were used for bacterial adhesion assays. Goats expressing HLZ in their milk were generated and express HLZ in milk at concentration of 270 μg/ml . Cells were incubated with pasteurized milk from either transgenic goats expressing HLZ or non-transgenic control goats in the presence and absence of EAEC strain 042 (O44:H18).

Results

Cellular proliferation was significantly greater in the presence of both HLZ transgenic and control goat milk compared to cells with no milk. Cellular migration was significantly decreased in the presence of EAEC alone but was restored in the presence of milk. Milk from HLZ transgenic goats had significantly more migration compared to control milk. Both milks significantly reduced EAEC adhesion to Caco-2 cells and transgenic milk resulted in less colonization than control milk using a HEp-2 assay. Both milks had significantly increased cellular viability as well as less apoptosis in both the absence and presence of EAEC.

Conclusions

These data demonstrated that goat milk is able to repair intestinal barrier function damage induced by EAEC and that goat milk with a higher concentration of lysozyme offers additional protection.

Keywords:
Intestinal cells; Cells proliferation; Cells migration; EAEC adherence; Cells viability; Apoptosis; Lysozyme