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

• Published in: BMC gastroenterology Vol. 12; no. 1; p. 106
• Main Authors: Carvalho, Eunice B, Maga, Elizabeth A, Quetz, Josiane S, Lima, Ila F N, Magalhães, Hemerson Y F, Rodrigues, Felipe A R, Silva, Antônio V A, Prata, Mara M G, Cavalcante, Paloma A, Havt, Alexandre, Bertolini, Marcelo, Bertolini, Luciana R, Lima, Aldo A M
• Format: Journal Article
• Language: English
• Published: England BioMed Central Ltd 11-08-2012; BioMed Central; BMC
• Subjects: Animal genetic engineering; Animals; Animals, Genetically Modified; Apoptosis; Apoptosis - drug effects; Bacteria; Bacterial Adhesion - drug effects; Bacterial infections; Cell Line, Tumor; Cell Movement - drug effects; Cell Proliferation - drug effects; Cell Survival - drug effects; Cells; Cells migration; Cells proliferation; Cells viability; Cells, Cultured; Colon cancer; Diarrhea; Dose-Response Relationship, Drug; EAEC adherence; Epithelium - drug effects; Epithelium - microbiology; Epithelium - pathology; Escherichia coli; Escherichia coli - drug effects; Escherichia coli - isolation & purification; Escherichia coli - physiology; Gastroenterology; Genetically modified animals; Goat's milk; Goats; Health aspects; Humans; In Vitro Techniques; Intestinal cells; Intestines - drug effects; Intestines - microbiology; Intestines - pathology; Lysozyme; Milk; Milk - enzymology; Muramidase - genetics; Muramidase - pharmacology; Physiological aspects; Proteins; Rats; Small intestine; Studies; Brazil; United States
• ISSN: 1471-230X; 1471-230X
• DOI: 10.1186/1471-230X-12-106

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.
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).
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.
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.