The Anti-inflammatory Effects of Glycerol-supplemented Probiotic Lactobacillus reuteri on Infected Epithelial cells In vitro

BACKGROUND: One of the most interesting effects of probiotics is their ability to modulate the immune system through the induction of cytokines and to enhance the host immune response. AIMS: The purpose of this study was to evaluate the anti-inflammatory effect of glycerol-supplemented Lactobacillus reuteri on the transcription level of interleukin (IL)-8 and human-beta-defensin (hBD)-2 expressed by epithelial cells after exposure to bacteria. MATERIALS AND METHODS: The confluent-cultured HaCat cell line (10(5) cells/mL) was exposed to Streptococcus mutans ATCC-25175 and Porphyromonas gingivalis ATCC-33277 (10(7) colony-forming units [CFU]/mL) for 24 h and challenged with probiotic L. reuteri ATCC-55730 (10(7) CFU/mL) supplemented with glycerol. Subsequently, the transcription levels of IL-8 and hBD-2 in HaCat cells were analyzed using reverse-transcription polymerase chain reaction (RT-PCR). In addition, cell viability was analyzed using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. All the obtained data were statistically analyzed using the one-way analysis of variance test, with P < 0.05 set as the level of significance. RESULTS: The MTT assays confirmed no cytotoxic effects of glycerol-supplemented L. reuteri on HaCat cells (viability >90%). mRNA expression of IL-8 and hBD-2 increased after exposure to both bacteria. The presence of glycerol-supplemented L. reuteri significantly reduced the expression of IL-8 and hBD-2 on HaCat cells (P < 0.05). CONCLUSION: Glycerol-supplemented L. reuteri reduced the expression of IL-8 and hBD-2, and the results may be proof of principle for a probiotic approach to combating inflammation. However, further studies are needed to validate this probiotic effect.