The effect of Bacillus coagulans on cytotoxicity and apoptosis induced by Salmonella Typhimurium in HT-29 cell culture

Abstract

Background and Objectives: Human epithelial cells have been widely used to study the interaction between intestinal cells and pathogens, in vitro. In this study, the effect of probiotic bacteria Bacillus coagulans and its supernatant on the growth inhibition, cytotoxicity and induction of apoptosis caused by Salmonella Typhimurium and its adhesion to HT-29 cells were investigated. Materials and Methods: B. coagulans supernatant was used to obtain the minimum inhibitory concentration. To evaluate the cytotoxicity and percent of apoptotic cells, B. coagulans and its supernatant (2, 4, 6 and 8% concentrations) with S. Typhimurium was added to HT-29 cells. The MTT assay was used in order to evaluate the cytotoxicity. Percent of apoptotic cells was reported using a fluorescence staining method. Additionally, the adhesion of S. Typhimurium to HT-29 cells was investigated. The effect of B. coagulans on the level of adhesion was also studied. Results: The most inhibitory effect was shown at the concentration of 80000 µg/ml supernatant of B. coagulans (54.77% ± 1.43). The simultaneous culture of S. Typhimurium with B. coagulans had the lowest amount of cytotoxicity and induced apoptosis among the all co-culture groups of S. Typhimurium with B. coagulans or its supernatant. The determined cytotoxicity and induced apoptosis were 26.06 % ± 3.79 and 17.63 % ± 2.14 respectively. In the adhesion test, it was observed that B. coagulans can significantly prevent adhesion of S. Typhimurium to HT-29 cell. Conclusion: B. coagulans can reduce the adhesion, cytotoxicity and induction of apoptosis caused by S. Typhimurium in HT-29 cells in vitro.

Keywords

• Probiotics
• Bacillus coagulans
• Fluorescence staining
• Minimum inhibitory concentration
• Salmonella Typh- imurium