title: Escherichia coli Nissle 1917-derived factors reduce cell death and late apoptosis and increase transepithelial electrical resistance in a model of 5-fluorouracil-induced intestinal epithelial cell damage.
abstract: We evaluated the capacity for supernatants (SNs) derived from Escherichia coli Nissle 1917 (EcN), cultured under different growth conditions, to prevent 5-fluorouracil (5-FU)-induced intestinal epithelial cell damage. EcN was cultured in: Luria Bertani (LB) broth, tryptone soya broth (TSB), de Man Rogosa Sharpe (MRS) broth, and M17 broth supplemented with 10% (v/v) lactose solution (M17). Intestinal epithelial cells (IEC-6) were treated with the following EcN SNs: LB(+), TSB(+), MRS(+), and M17(+) in the presence and absence of 5-FU (1.5 or 5 μM). Cell viability, apoptotic activity and cell monolayer permeability were measured by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT), flow cytometry, and transepithelial electrical resistance (TER) assays, respectively. 5-FU significantly reduced cell viability (P<0.05) at both 24 and 48 h. However, only EcN SN produced from LB and M17 growth media significantly decreased cell death induced by 5-FU (by approximately 10% after 24 and 48 h; and 10% after 24 h, respectively [P<0.05]). When measured by flow cytometry all EcN SNs in the presence of 5-FU increased the proportion of viable cells (by 3-5% for 24 h, 3-7% for 48 h, P<0.05) and reduced late-apoptotic cells after 24 and 48 h, compared with 5-FU control. Moreover, all EcN SNs significantly reduced the disruption of IEC-6 cell barrier function induced by 5-FU by 7-10% (P<0.05), compared with DMEM control. We conclude that EcN derived factors could potentially reduce the severity of intestinal mucositis.
authors.affiliation: School of Animal and Veterinary Sciences; University of Adelaide; Roseworthy, SA Australia.
authors.forename: Hanru
authors.initials: H
authors.lastname: Wang
authors.affiliation: School of Agriculture, Food and Wine; University of Adelaide; Waite Campus; Urrbrae, SA Australia.
authors.forename: Susan E P
authors.initials: SE
authors.lastname: Bastian
authors.affiliation: Gastroenterology Department; Women's and Children's Hospital; Adelaide, SA Australia.
authors.forename: Ker Y
authors.initials: KY
authors.lastname: Cheah
authors.affiliation: Microbiology Department; SA Pathology at Women's and Children's Hospital; Adelaide, SA Australia.
authors.forename: Andrew
authors.initials: A
authors.lastname: Lawrence
authors.affiliation: School of Animal and Veterinary Sciences; University of Adelaide; Roseworthy, SA Australia; Gastroenterology Department; Women's and Children's Hospital; Adelaide, SA Australia.
authors.forename: Gordon S
authors.initials: GS
authors.lastname: Howarth
mesh_terms: D000818:Animals; D000964:Antimetabolites, Antineoplastic / Q000009:adverse effects*; D017209:Apoptosis / Q000187:drug effects; D001688:Biological Products / Q000302:isolation & purification / Q000494:pharmacology*; D016923:Cell Death; D002460:Cell Line; D002470:Cell Survival / Q000187:drug effects; D017077:Culture Media, Conditioned / Q000737:chemistry; D017097:Electric Impedance; D004847:Epithelial Cells / Q000187:drug effects* / Q000473:pathology; D004926:Escherichia coli / Q000737:chemistry*; D005472:Fluorouracil / Q000009:adverse effects*; D007082:Ileum / Q000187:drug effects* / Q000473:pathology; D009336:Necrosis / Q000473:pathology; D010539:Permeability; D051381:Rats
chemical_list: D000964:Antimetabolites, Antineoplastic; D001688:Biological Products; D017077:Culture Media, Conditioned; D005472:Fluorouracil
keywords: IEC-6 cells; chemotherapy; cytotoxicity; mucositis