Cytotoxicity, apoptosis, and IL-8 gene expression induced by some foodborne pathogens in presence of Bacillus coagulans in HT-29 cells

Abstract

Food poisoning caused by bacteria is one of the most important concerns in food hygiene. The use of probiotics in prevention, control, and treatment of these infections has been considerably increased in recent years. This study evaluated the effect of B. coagulans cell free supernatant (CFS) on growth of Bacillus cereus, Listeria monocytogenes, Staphylococcus aureus, non-pathogenic Escherichia coli, and Escherichia coli 0157:H7 by the broth dilution method. The cytotoxicity, and apoptosis induced by pathogens alone and in co-culture with B. coagulans or its CFS were measured by trypan blue, and fluorescence staining methods. The expression level of interleukin-8 (IL-8) cytokine-encoding genes was also investigated by a qRT-PCR assay in all pathogens and co-cultured groups in HT-29 cells. Our results showed that 4% B. coagulans CFS reduced pathogen growth. The highest rate of growth inhibition was observed in L. monocytogenes. We also found that B. coagulans, and its 4% CFS reduced the cytotoxic effects of pathogens, with the exception of S. aureus. Non-pathogenic E. coli also had no significant cytotoxic effect on the cells. Examination of the treated cells with acridine orange/ethidium bromide staining showed reductions in the rate of cell damage (including early apoptosis, late apoptosis, and necrosis) in pathogen-probiotic co-cultures. Furthermore, we showed that co-culture of pathogens with B. coagulans significantly down-regulated IL-8 gene expression (P < 0.05). The greatest down-regulation compared with pathogen alone was observed in S. aureus. Hence, B. coagulans can be considered as an appropriate probiotic to diminish cytotoxicity, and inflammatory response of enteropathogenic bacteria.