title: Inhibition of Lipopolysaccharide-Induced Interleukin 8 in Human Adenocarcinoma Cell Line HT-29 by Spore Probiotics: B. coagulans and B. subtilis (natto).
abstract: Probiotics are used as a treatment for different intestinal disorders. They confer health benefits by different ways. This study was aimed to investigate immunomodulatory effect of Bacillus probiotic spores on the production of lipopolysaccharide (LPS)-induced interleukin 8 (IL-8) in HT-29 intestinal epithelial cells. Differentiated intestinal epithelial cell line was used as a model for the study of colonization of purified spores (Bacillus subtilis (natto) and B. coagulans) and their anti-inflammatory effects. MTT assay and trypan blue staining were used for the detection of optimal concentration of the purified spores and LPS. Pre-treatment assay was done by treatment of the cells with the purified spores for 2 h, followed by challenges with LPS for 3 and 18 h. Post-treatment assay was done by initial treatment of the cells with LPS for 18 h, followed by the spores for 3 and 6 h. Levels of IL-8 secretion and its mRNA expression were measured by ELISA and relative Q real-time PCR. Our results showed similar rates of adherence to intestinal epithelial cells by the spore probiotics, while displaying no cytotoxic effect. In the pre-treatment assay, a significant decrease in IL-8, at both protein and mRNA levels, was measured for B. coagulans spores after the addition of LPS, which was higher than those observed for Bacillus subtilis (natto) spores. In the post-treatment assay, while Bacillus subtilis (but not B. coagulans) diminished the LPS-stimulated IL-8 levels after 3 h of incubation, the inhibitory effect was not constant. In conclusion, ability of Bacillus spore probiotics for adherence to intestinal epithelial cell and their anti-inflammatory effects, through interference with LPS/IL-8 signaling, was shown in this study. Further studies are needed to characterize responsible bacterial compounds associated with these effects.
authors: Foodborne and Waterborne Diseases Research Center, Research Institute for Gastroenterology and Liver Diseases, Shahid Beheshti University of Medical Sciences, Tehran, Iran.|Department of Cell and Molecular Biology, Pharmaceutical Sciences Branch, Islamic Azad University, Tehran, Iran.
authors: Masoumeh
authors: M
authors: Azimirad
authors: Foodborne and Waterborne Diseases Research Center, Research Institute for Gastroenterology and Liver Diseases, Shahid Beheshti University of Medical Sciences, Tehran, Iran. masoud.alebouyeh@gmail.com.|Gastroenterology and Liver Diseases Research Center, Research Institute for Gastroenterology and Liver Diseases, Shahid Beheshti University of Medical Sciences, Tehran, Iran. masoud.alebouyeh@gmail.com.
authors: Masoud
authors: M
authors: Alebouyeh
authors: Department of Cell and Molecular Biology, Pharmaceutical Sciences Branch, Islamic Azad University, Tehran, Iran.
authors: Tahereh
authors: T
authors: Naji
mesh_terms: D000893:Anti-Inflammatory Agents / Q000494:pharmacology*; D001407:Bacillus / Q000502:physiology*; D001412:Bacillus subtilis / Q000502:physiology*; D045744:Cell Line, Tumor; D019073:HT29 Cells; D006801:Humans; D016209:Interleukin-8 / Q000235:genetics* / Q000276:immunology; D007422:Intestines / Q000187:drug effects / Q000276:immunology / Q000382:microbiology; D008070:Lipopolysaccharides / Q000009:adverse effects / Q000276:immunology; D019936:Probiotics / Q000494:pharmacology*; D013171:Spores, Bacterial / Q000502:physiology
chemical_list: D000893:Anti-Inflammatory Agents; D016209:Interleukin-8; D008070:Lipopolysaccharides
keywords: Bacillus coagulans; Bacillus subtilis (natto); Immunomodulatory effect; Lipopolysaccharide