Ameliorating inflammation in an in vitro model by screening the anti-inflammatory and immunomodulatory roles of putative probiotics in inflammatory bowel disease.

IBD is considered a relapsing disease with relapsing phases. Probiotics are beneficial microorganisms that modulate inflammatory signaling pathways. Our aim was to identify the precise molecular effects of probiotics on inflammatory signaling pathways during the presence of inflammation. Evaluation of the expression of JAK/STAT and inflammatory genes after treatment of the HT -29 cell line with the sonicated pathogens and probiotics, simultaneously was performed by quantitative real-time polymerase chain reaction (qPCR) assay. The production of IL-6 and IL-1ß after administration of probiotics was conducted by means of cytokine assay. The probiotic cocktail resulted in the downregulation of TIRAP, IRAK4, NEMO, and RIP genes in the NF-кB pathway compared with Sonicat-treated cells. The expression of JAK/STAT genes was various after probiotic treatment. The application of probiotics has been observed to result in a notable decrease in the production of IL-6 and IL-1ß. The investigated probiotic cocktail, especially Bifidobacterium spp. showed anti-inflammatory effects on HT -29 cells via modulation of JAK/STAT and NF-кB signaling pathways. The use of probiotics with the least side effects could be considered a suitable treatment for patients with inflammatory bowel disease, even at the beginning of inflammation.

Investigation of the anti-inflammatory effects of native potential probiotics as supplementary therapeutic agents in an in-vitro model of inflammation.

BACKGROUND: IBD is considered an inflammatory disease with abnormal and exaggerated immune responses. To control the symptoms, different theraputic agents could be used, however, utilizing the agents with the least side effects could be important. Probiotics as beneficial microorganisms are one of the complementory theraputic agents that could be used to modulate inflammatory signaling pathways. In the current study, we aimed to identify the precise molecular effects of potential probiotics on signaling pathways involved in the development of inflammation. METHODS: A quantitative real-time polymerase chain reaction (qPCR) assay was used to analyze the expression of JAK /STAT (JAK1, JAK2, JAK3, TYK2, STAT1, STAT2, STAT3, STAT4, STAT5 and STAT6) and inflammatory genes (NEMO, TIRAP, IRAK, and RIP) after the HT -29 cell line treatment with the sonicated pathogens and potential probiotics. A cytokine assay was also used to evaluate IL -6 and IL -1ß production after potential probiotic treatment. RESULTS: The potential probiotic cocktail downregulated the JAK genes and TIRAP, IRAK4, NEMO, and RIP genes in the NF-kB pathway compared with cells that were treated with sonicated gram negative pathogens. The expression of STAT genes was different after potential probiotic treatment. The production of IL -6 and IL -1ß decreased after potential probiotic treatment. CONCLUSIONS: Considering the importance of controlling the symptoms of IBD to improve the life quality of the patients, using probiotic could be crucial. In the current study the studied native potential probiotic cocktails showed anti-inflammatory effects via modulation of JAK /STAT and NF-kB signaling pathways. This observation suggests that our native potential probiotics consumption could be useful in reducing intestinal inflammation.

The Role of Combining Probiotics in Preventing and Controlling Inflammation: A Focus on the Anti-Inflammatory and Immunomodulatory Effects of Probiotics in an In Vitro Model of IBD.

Objective: IBD is an inflammatory disease with abnormalities such as dysbiosis and abnormal immune system activity. Probiotics, as live beneficial microorganisms, play a role in maintaining health through various mechanisms, including the modulation of the immune system and the control of inflammation. Here, we aimed to investigate the efficacy of a probiotic mixture of Lactobacillus spp. and Bifidobacterium spp. in modulating JAK/STAT and NF-kB inflammatory signaling pathways. Method: A quantitative real-time polymerase chain reaction (qPCR) assay was conducted to analyze the expression of JAK/STAT and inflammatory genes after treatment with the probiotic mixture before, after, and simultaneously with the sonicated pathogen in the HT-29 cell line. The production of IL-6 and IL-1ß after probiotic treatment was investigated via cytokine assay. Results: Treatment with probiotics resulted in downregulation of TIRAP, IRAK4, NEMO, and RIP genes in the NF-kB pathway and JAK/STAT genes compared with sonicat-treated cells as inflammation inducers. The production of IL-6 and IL-1 decreased after probiotic treatment. Conclusions: The probiotic mixture of Lactobacillus spp. and Bifidobacterium spp. showed anti-inflammatory effects by modulating JAK/STAT and NF-kB signaling pathways. The use of probiotics could be considered as an appropriate complementary treatment for patients with inflammatory bowel disease.

The impact of Lactobacillus and Bifidobacterium probiotic cocktail on modulation of gene expression of gap junctions dysregulated by intestinal pathogens.

Probiotics are special bacterial strains with strain specific impacts. They can affect health condition in intestine by producing organic acid, competing with pathogens and maintaining cells homeostasis. Regarding to importance of cell junctions in cells transportation and the influence of pathogens in their functions which lead to inflammation, the impact of probiotic strains comprised of Lactobacillus and Bifidobacterium strains on two important members of gap junctions (Cx26 and Cx43) were assayed. The expressions of cell junction genes in contact with probiotic cocktail along with pathogenic components of enterotoxigenic Escherichia coli and Salmonella typhimurium on HT-29 cell line in different treatment orders were evaluated. Results analysis demonstrated downregulation of cx26 and cx43 along with pathogenic components while, probiotic cocktail could modulate their expression by upregulation. We concluded that Lactobacillus and Bifidobacterium strains were efficient probiotics, when they were used as one cocktail, impacted grater amount on the expression of cell junctions and this might lead to modulate homeostasis and reveal inflammation symptoms in intestine.

Comparison of the prevalence of bacteriocin encoding genes in Lactobacillus spp. isolated from fecal samples of healthy volunteers, IBD-patient and IBD-recovered.

Background and Objectives: Bacteriocins are antimicrobial peptides produced by many genera of bacteria especially Lactobacillus spp. against many pathogens, adapt bacterial composition in the gut and inhibit dysbiosis that can lead to inflammation disorders like inflammatory bowel disease (IBD). The aim of this study was to compare the prevalence of bacteriocin genes in health, IBD disease and recovery conditions. Materials and Methods: In this survey 115 Lactobacillus spp. from 58 fecal samples of three different groups were evaluated. Comparison of the presence of bacteriocin genes in different groups were assayed by purified samples and PCR method, followed by statistical analysis to identify the effect of inflammation in the proportion of Lactobacillus spp. and presence of their bacteriocin genomes. Results: Of 115 Lactobacillus spp. 60% of samples had positive bacteriocin-encoding genes which included: gassericin-A 29.56%, acidocin 15.65%, plantaricin-NC8 18.26%, plantaricin-S 13.04%, lactacin-F 9.5%, sakacin-P 6.08% and gassericin-T 6.08%. Results indicated that the percentage of positive bacteriocin genes were much more in healthy volunteer and IBD-recovered in comparison to IBD-patients which showed the effect of inflammation in the presence of bacteriocin genes. Conclusion: The results obtained in this study demonstrated that the presence of bacteriocin genes can be related to health and disease states and inflammatory disease affected the prevalence of bacteriocin-encoding genes. This approach can help to identify bacterial functions that can be targeted in future concepts of IBD therapy.

Anti-inflammatory and immunomodulatory effects of Lactobacillus spp. as a preservative and therapeutic agent for IBD control.

BACKGROUND: Probiotics have a beneficial effect on inflammatory responses and immune regulation, via Janus kinase/signal transduction and activator of transcription (JAK/STAT) and NF-κB signaling pathways. To evaluate the precise effects of Lactobacillus spp. as a protective and therapeutic agent, we aimed to investigate the efficacy of Lactobacillus spp. in modulating JAK/STAT and nuclear factor kappa B (NF-κB) inflammatory signaling pathways. METHODS: A quantitative real-time polymerase chain reaction (qPCR) assay was used to analyze the expression of JAK/STAT and inflammatory genes (TIR-associated Protein [TIRAP], Interleukin 1 Receptor Associated Kinase[IRAK4], Nuclear factor-kappa B Essential Modulator [NEMO], and receptor interacting protein [RIP]) followed by treatment of the HT-29 cell line with sonicated pathogens before, after, and simultaneously with Lactobacillus spp. A cytokine assay was also used to evaluate interleukin (IL)-6 and IL-1ß production after treatment with Lactobacillus spp. RESULTS: Lactobacillus spp. downregulated JAK and TIRAP, IRAK4, NEMO, and RIP genes in the NF-κB pathway compared to sonicate-treated cells. The expression of STAT genes was different after treatment with probiotics. The production of IL-6 and IL-1ß decreased after probiotic treatment. CONCLUSIONS: Our Lactobacillus spp. cocktail showed anti-inflammatory effects on HT-29 cells by modulating JAK/STAT and NF-κB signaling pathways in all three treatment variants. Therefore, Lactobacillus spp. as a dietary supplement can both prevent and reduce inflammation-related diseases such as inflammatory bowel disease.

The effects of the probiotic cocktail on modulation of the NF-kB and JAK/STAT signaling pathways involved in the inflammatory response in bowel disease model.

BACKGROUND: Probiotics positively affect inflammatory responses, in part, through Janus kinase/signal transduction and activator of transcription (JAK/STAT) and inflammatory signaling pathways. To evaluate the precise effects of probiotics as protective treatment, we aimed to investigate the effectiveness of Lactobacillus spp., Bifidobacterium spp., and a mixture of these probiotics in modulating the JAK/STAT and inflammatory signaling pathways. METHODS: A quantitative real-time polymerase chain reaction (qPCR) assay was used to analyze the expression of JAK/STAT and inflammatory genes (TIRAP, IRAK4, NEMO, and RIP) following HT-29 cell line treatment with sonicated pathogens Lactobacillus spp., Bifidobacterium spp., and a mixed cocktail. A cytokine assay was also used to evaluate the IL-6 and IL-1ß production following the probiotic treatment. RESULTS: The probiotic cocktail downregulated the JAK genes and TIRAP, IRAK4, NEMO, and RIP genes in the NF-kB pathway compared to sonicate pathogen treatment cells. The expression of STAT genes was variable following probiotic treatment. The IL-6 and IL-1ß production decreased after probiotic treatment. CONCLUSIONS: Our probiotic cocktail showed anti-inflammatory effects on HT-29 cells by modulating JAK/STAT and NF-kB pathways. Therefore, Lactobacillus spp. and Bifidobacterium spp. probiotics as nutritional supplements may reduce inflammation-associated diseases such as inflammatory bowel disease (IBD).

Identification of different Escherichia coli pathotypes in north and north-west provinces of Iran.

BACKGROUND AND OBJECTIVES: Diarrhea is one of the most prevalent diseases in the world, specially in developing countries. One of the most important causative agents of bacterial diarrhea is diarrheagenic Escherichia coli (DEC) which causes gastroenteritis and this group involving enterotoxigenic E. coli (ETEC), enteropathogenic E. coli (EPEC), enteroaggregative E. coli (EAEC), enterohemoragic E. coli (EHEC), enteroinvasive E. coli (EIEC), diffusely adherence E. coli (DAEC). The aim of this study was to identify different E. coli pathotypes in north and north-west of Iran, among the clinical isolates. MATERIALS AND METHODS: In this study for identification of E. coli, 170 fecal samples were cultured on MacConkey agar and identified by biochemical tests. Samples with E. coli characteristics were selected (145 samples) and their genomes were purified by phenol-chloroform method. After extraction of genomes, lt and sta genes identified by PCR for ETEC, eae gene for atypical and eae and bfp for typical EPEC, AA region for EAEC, stx1 and stx2 and eae genes for EHEC (stx1 or stx2 or both for STEC) and invE for EIEC. RESULTS: Finally 10 samples identified as ETEC (%5.88), 18 (%10.58) EPEC, 6 (%3.52) EHEC and 12 (7.05%) samples were STEC. None of the samples were positive for EAEC and EIEC. CONCLUSION: The results obtained in this study showed that ETEC, EPEC, EHEC and STEC are prevalent bacterial agents in north and north-west of Iran. Complementary studies to identify these pathotypes in other seasons can help to adopt necessary policies against outbreaks in Iran.