Lactobacillus plantarum L15 Alleviates Colitis by Inhibiting LPS-Mediated NF-?B Activation and Ameliorates DSS-Induced Gut Microbiota Dysbiosis.
ABSTRACT: Previous studies have suggested that the Lactobacillus plantarum bacteria strain could be effective in ulcerative colitis (UC) management. However, its effects are strain-specific and the related mechanisms for its attenuating effects on UC remain unclear. This study aimed to elucidate the underlying mechanisms for the protective effect of L. plantarum on UC. Firstly, 15 L. plantarum strains were screened for potential probiotic characteristics with good tolerance to simulated human gastrointestinal transit and adhesion. Secondly, the inflammatory response of selected strains to the Caco-2 cells induced by lipopolysaccharide (LPS) was measured. Finally, an in vivo mouse model induced by dextran sulfate sodium (DSS) was used to assess the beneficial effects and likely action mechanisms the successfully screened in vitro strain, L. plantarum L15. In vitro results showed that L. plantarum L15 possessed the highest gastrointestinal transit tolerance, adhesion and reduction of pro-inflammatory abilities compared to the other screened strains. In vivo, high dose of L. plantarum L15 supplementation increased the body weight, colon length and anti-inflammatory cytokine production. Pro-inflammatory cytokine production, disease activity index (DAI) levels and myeloperoxidase (MPO) parameters decreased using this strain. In addition, L. plantarum L15 alleviated the histopathological changes in colon, modulated the gut microbiota, and decreased LPS secretion. The activities of this strain down-regulated the expression of TLR4 and MyD88 genes as well as genes associated with NF-?B signaling pathway. Our findings present L. plantarum L15 as a new probiotic, with promising application for UC management.
Project description:The present study evaluates the probiotic properties of three Lactobacillus plantarum strains MJM60319, MJM60298, and MJM60399 possessing antimicrobial activity against animal enteric pathogens. The three strains did not show bioamine production, mucinolytic and hemolytic activity and were susceptible to common antibiotics. The L. plantarum strains survived well in the simulated orogastrointestinal transit condition and showed adherence to Caco-2 cells in vitro. The L. plantarum strains showed strong antimicrobial activity against enterotoxigenic Escherichia coli, Shiga toxin-producing E. coli, Salmonella enterica subsp. enterica serovar Typhimurium, Choleraesuis and Gallinarum compared to the commercial probiotic strain Lactobacillus rhamnosus GG. The mechanism of antimicrobial activity of the L. plantarum strains appeared to be by the production of lactic acid. Furthermore, the L. plantarum strains tolerated freeze-drying and maintained higher viability in the presence of cryoprotectants than without cryoprotectants. Finally, the three L. plantarum strains tolerated NaCl up to 8% and maintained >60% growth. These characteristics of the three L. plantarum strains indicate that they could be applied as animal probiotic after appropriate in vivo studies.
Project description:<i>Lactiplantibacillus plantarum</i> are probiotic bacteria, maintaining the integrity of the gastrointestinal epithelial barrier, and preventing the infection of pathogenic bacteria. Exopolysaccharides (EPSs) are often involved in the probiotic property of <i>L. plantarum</i>. Here, we identified a new EPS-synthetic gene cluster, <i>cpsWc</i>, carrying 13 genes, laid on a large plasmid in a well-characterized probiotic <i>L. plantarum</i> strain LTC-113. The <i>cpsWc</i> gene cluster was horizontally acquired from <i>Weissella cibaria</i>, enhancing the biofilm formation ability of the host strain and its tolerance to harsh environmental stresses, including heat, acid, and bile. Transfer of <i>cpsWc</i> also conferred the probiotic properties to other <i>L. plantarum</i> strains. Moreover, <i>cpsWc</i> strengthened the adhesion of LTC-113 to intestinal epithelial cells. Both the <i>cpsWc</i>-carrying LTC-113 and its EPSs <i>per se</i> effectively attenuated the LPS-induced pro-inflammatory effect of intestinal epithelial cells, and inhibited the adhesion of pathogenic bacteria, such as <i>S. typhimurium</i> and <i>E. coli</i> by exclusion and competition. The newly identified <i>cpsWc</i> gene cluster emphasized the contribution of mobile EPS-synthetic element on the probiotic activity of <i>L. plantarum</i>, and shed a light on the engineering of probiotic bacteria.
Project description:Three Lactobacillus plantarum strains ATG-K2, ATG-K6, and ATG-K8 were isolated from Kimchi, a Korean traditional fermented food, and their probiotic potentials were examined. All three strains were free of antibiotic resistance, hemolysis, and biogenic amine production and therefore assumed to be safe, as supported by whole genome analyses. These strains demonstrated several basic probiotic functions including a wide range of antibacterial activity, bile salt hydrolase activity, hydrogen peroxide production, and heat resistance at 70°C for 60 s. Further studies of antimicrobial activities against Candida albicans and Gardnerella vaginalis revealed growth inhibitory effects from culture supernatants, coaggregation effects, and killing effects of the three probiotic strains, with better efficacy toward C. albicans. In vitro treatment of bacterial lysates of the probiotic strains to the RAW264.7 murine macrophage cell line resulted in innate immunity enhancement via IL-6 and TNF-? production without lipopolysaccharide (LPS) treatment and anti-inflammatory effects via significantly increased production of IL-10 when co-treated with LPS. However, the degree of probiotic effect was different for each strain as the highest TNF-? and the lowest IL-10 production by the RAW264.7 cell were observed in the K8 lysate treated group compared to the K2 and K6 lysate treated groups, which may be related to genomic differences such as chromosome size (K2: 3,034,884 bp, K6: 3,205,672 bp, K8: 3,221,272 bp), plasmid numbers (K2: 3, K6 and K8: 1), or total gene numbers (K2: 3,114, K6: 3,178, K8: 3,186). Although more correlative inspections to connect genomic information and biological functions are needed, genomic analyses of the three strains revealed distinct genomic compositions of each strain. Also, this finding suggests genome level analysis may be required to accurately identify microorganisms. Nevertheless, L. plantarum ATG-K2, ATG-K6, and ATG-K8 demonstrated their potential as probiotics for mucosal health improvement in both microbial and immunological contexts.
Project description:<b>Background</b>: Probiotic bacteria with anti-inflammatory properties have the potential to be of therapeutic benefit in gingivitis. <b>Objective</b>: To evaluate the effects of potential probiotic strains on inflammatory mediators involved in early gingivitis using an <i>ex vivo</i> inflammation model. <b>Methods</b>: Strains were screened in viable and attenuated forms for effects on bacterial lipopolysaccharide (LPS)-stimulated release of interleukins (IL)-1?, -6 and -8, tumor necrosis factor-?, prostaglandin E<sub>2</sub> and 8-isoprostane from human primary monocytes, and then, if anti-inflammatory effects were shown, on IL-1?-stimulated release of inflammatory mediators from primary gingival fibroblasts. Lead strains were evaluated for optimal dosing, batch-to-batch variation and functional consistency in toothpaste. <b>Results</b>: Twenty-one of 73 strains showed anti-inflammatory effects in monocytes; of which, seven showed effects in both viable and attenuated forms. Seven of 14 strains showed effects in fibroblasts. Strains <i>Lactobacillus paracasei</i> LPc-G110(SYBIO-15) and <i>Lactobacillus plantarum</i> GOS42(SYBIO-41) induced statistically significant dose-dependent reductions in the release of multiple inflammatory mediators from monocytes, which were consistent across batches. Viable <i>L. paracasei</i> LPc-G110 tooth paste significantly reduced IL-6, IL-8 and prostaglandin E<sub>2</sub> release from monocytes versus placebo. <b>Conclusion</b>: Strains <i>L. paracasei</i> LPc-G110 and <i>L. plantarum</i> GOS42 have potential for use as probiotics in oral care products to reduce gingival inflammation.
Project description:<h4>Aim</h4>To analyze the changes of different Lactobacillus species in ulcerative colitis patients and to further assess the therapeutic effects of selected Lactobacillus strains on dextran sulfate sodium (DSS)-induced experimental colitis in BALB/c mice.<h4>Methods</h4>Forty-five active ulcerative colitis (UC) patients and 45 population-based healthy controls were enrolled. Polymerase chain reaction (PCR) amplification and real-time PCR were performed for qualitative and quantitative analyses, respectively, of the Lactobacillus species in UC patients. Three Lactobacillus strains from three species were selected to assess the therapeutic effects on experimental colitis. Sixty 8-week-old BALB/c mice were divided into six groups. The five groups that had received DSS were administered normal saline, mesalazine, L. fermentum CCTCC M206110 strain, L. crispatus CCTCC M206119 strain, or L. plantarum NCIMB8826 strain. We assessed the severity of colitis based on disease activity index (DAI), body weight loss, colon length, and histologic damage.<h4>Results</h4>The detection rate of four of the 11 Lactobacillus species decreased significantly (P < 0.05), and the detection rate of two of the 11 Lactobacillus species increased significantly (P < 0.05) in UC patients. Relative quantitative analysis revealed that eight Lactobacillus species declined significantly in UC patients (P < 0.05), while three Lactobacillus species increased significantly (P < 0.05). The CCTCC M206110 treatment group had less weight loss and colon length shortening, lower DAI scores, and lower histologic scores (P < 0.05), while the CCTCC M206119 treatment group had greater weight loss and colon length shortening, higher histologic scores, and more severe inflammatory infiltration (P < 0.05). NCIMB8826 improved weight loss and colon length shortening (P < 0.05) with no significant influence on DAI and histologic damage in the colitis model.<h4>Conclusions</h4>Administration of an L. crispatus CCTCC M206119 supplement aggravated DSS-induced colitis. L. fermentum CCTCC M206110 proved to be effective at attenuating DSS-induced colitis. The potential probiotic effect of L. plantarum NCIMB8826 on UC has yet to be assessed.
Project description:The anti-inflammatory potential of eight indigenous probiotic Lactobacillus isolates was evaluated in vitro in terms of modulating the expression of tumor necrosis factor-alpha (TNF-?) and interleukin-6 (IL-6) in human acute monocytic leukemia (THP-1) cells under inflammatory conditions. Amongst these, Lactobacillus plantarum Lp91 was the most potent anti-inflammatory strain as it evoked a significant (P < 0.001) down-regulation of TNF-? by -1.45-fold relative to the control in THP-1 cells. However, in terms of IL-6 expression, all the strains could up-regulate its expression considerably at different levels. Hence, based on in vitro expression of TNF-?, Lp91 was selected for in vivo study in lipopolysaccharide (LPS)-induced mouse model to look at the expression of TNF-?, IL-6, monocyte chemotactic protein-1 (MCP-1), vascular cell adhesion molecule-1 (VCAM-1), intercellular adhesion molecule (ICAM-1) and E-selectin in mouse aorta. In LPS challenged (2 h) mice group fed with Lp91 for 10 days, TNF-?, IL-6, MCP-1, VCAM-1, ICAM-1 and E-selectin expressions were significantly down-regulated by 3.10-, 10.02-, 4.22-, -3.14-, 2.28- and 5.71-fold relative to control conditions. In conclusion, Lp91 could serve as a candidate probiotic strain to explore it as a possible biotherapeutic anti-inflammatory agent against inflammatory diseases including cardiovascular disease.
Project description:This study aimed to evaluate the safety (hemolysis and enzyme activity), probiotic properties (gastrointestinal tract tolerance, adhesion, hydrophobicity, and auto-aggregation), and functional characteristics (antimicrobial, antioxidant, and ?-galactosidase activities) of lactic acid bacteria (LAB), isolated from kimchi, in order to select a multifunctional LAB strain for starter culture in fermented food. The five isolated strains included Lactobacillus plantarum WiKim83, L. plantarum WiKim84, Pediococcus pentosaceus WiKim85, P. pentosaceus WiKim86, and L. plantarum WiKim87, as identified by 16S rRNA gene sequence analysis; they were confirmed to be nonhemolytic and not able to produce ?-glucuronidase, a carcinogenic enzyme. Probiotic properties of the five LAB strains were evaluated relative to those of commercial Lactobacillus rhamnosus GG, and results revealed probiotic potential of three strains (L. plantarum WiKim83, L. plantarum WiKim84, and L. plantarum WiKim87) to be superior. L. plantarum WiKim84 showed high antimicrobial activity against pathogens, and L. plantarum WiKim83 exhibited the highest antioxidant and ?-galactosidase activities. Based on the probiotic and functional properties, the main characteristics of each strain were highlighted and two of them, L. plantarum WiKim83 and L. plantarum WiKim87, were selected as the most potent by principal component analysis. These strains showed antimicrobial, ?-galactosidase, and antioxidant activities, which recommend their suitability as starter culture in various fermented foods.
Project description:Fermented products, including <i>Ogi-baba</i> and <i>Pito</i>, provide several health benefits, particularly when probiotics are used in the fermentation process. Probiotic microorganisms exert strain-specific health-promoting activities on humans and animals. The objective of this study was to investigate the probiotic potentials of Lactic-acid bacteria (LAB) isolates from indigenous fermented sorghum products (<i>Ogi-baba</i> and <i>Pito</i>). The LAB isolates were screened for potential probiotic properties by antagonistic activity against eight enteropathogenic clinical bacteria isolates (<i>Escherichia coli, Klebsiella</i> sp.<i>, Helicobacter pylori, Bacillus</i> sp.<i>, Staphylococcus</i> sp.<i>, Salmonella</i> sp.<i>, Pseudomonas</i> sp. and <i>Listeria monocytogenes</i>) as indicator organisms using the agar well diffusion technique. The organisms were also screened for acidity, bile tolerance, antibiotic susceptibility, production of lactic acid, diacetyl and hydrogen peroxide. β-galactosidase assay was also done. Genomic DNA was extracted from the two selected LAB isolates; the 16S rRNA were amplified and sequenced. The sequence data were subjected to Basic Local Alignment Search Tool (BLAST) and molecular phylogenetic analyses to identify the isolates. The isolates were identified as strains of <i>Lactobacillus plantarum</i> and <i>Pediococcus pentosaceus.</i> The sequence data for these two isolates were submitted to the Genbank with accession numbers KP883298 and KP883297 respectively. The <i>P. pentosaceus</i> strain (PB2) strain exhibited β-galactosidase activity as well as <i>L. plantrum</i> strain (OB6). The study revealed exceptional probiotic potentials of two LAB namely <i>Lactobacillus plantarum</i> strain (OB6) and <i>Pediococcus pentosaceus</i> strain (PB2) isolated from fermented sorghum products, <i>Ogi-baba</i> and <i>Pito</i> respectively. Hence, the two LAB strains may be potentially used as probiotic to prevent some enteropathogen-induced gastrointestinal disorders; reduce the incidence of respiratory tract infections and for the management of lactose in intolerance.
Project description:In this study, Lactobacillus plantarum strain DHCU70 isolated from dahi, a fermented milk product and L. plantarum strain DKP1 isolated from kinema, a fermented soybean food of India, respectively were evaluated for their bacteriocin production and probiotic properties. Both strains of L. plantarum (DHCU70 and DKP1) were found to have potent antimicrobial activity against Kocuria rhizophila ATCC 9341. Bacteriocin produced by L. plantarum strains DHCU70 and DKP1 did not exhibit inhibition of cell wall, DNA and fatty acids biosynthesis mechanisms as evaluated by whole cell reporter assays. We characterized the bacteriocin encoding genes in L. plantarum strains DHCU70 and DKP1 by whole genome sequence which consisted of a single and circular chromosome with genome size of 3.38 Mb (GC content of 44.3%) and 3.39 Mb, respectively and a GC content of 44.3%. L. plantarum DHCU70 has 3252 number of protein encoding genes comprising 89 number of RNA genes (69tRNA, 16rRNA, 4nc RNA) whereas L. plantarum DKP1 has total of 3277 number of protein encoding genes with 89 number. of RNA genes (69tRNA, 16S rRNA, 4nc RNA). Analysis revealed the presence of 20.5 kb long and 23 numbers of plantaricin encoding locus (pln locus) for production of antimicrobial compound. BAGEL analysis has shown that the pln locus of both the strains of L. plantarum showed maximum sequence similarity with plantaricin NC8 of L. plantarum NC8, originally isolated from grass silage. Annotated whole genome sequence of both strains DHCU70 and DKP1 was analyzed for the presence of probiotic marker genes. The probiotic properties of these strains of were also evaluated in vitro. Due to the presence of genes responsible for antimicrobial activity and probiotic properties, both strains of L. plantarum may be considered as a suitable probiotic candidate in food industry.
Project description:BACKGROUND:We evaluated the functional capacity of plantaricin-producing Lactobacillus plantarum SF9C and S-layer-carrying Lactobacillus brevis SF9B to withstand gastrointestinal transit and to compete among the gut microbiota in vivo. Considering the probiotic potential of Lb. brevis SF9B, this study aims to investigate the antibacterial activity of Lb. plantarum SF9C and their potential for in vivo colonisation in rats, which could be the basis for the investigation of their synergistic functionality. RESULTS:A plantaricin-encoding cluster was identified in Lb. plantarum SF9C, a strain which efficiently inhibited the growth of Listeria monocytogenes ATCC® 19111™ and Staphylococcus aureus 3048. Homology-based three-dimensional (3D) structures of SF9C plantaricins PlnJK and PlnEF were predicted using SWISS-MODEL workspace and the helical wheel representations of the plantaricin peptide helices were generated by HELIQUEST. Contrary to the plantaricin-producing SF9C strain, the S-layer-carrying SF9B strain excluded Escherichia coli 3014 and Salmonella enterica serovar Typhimurium FP1 from the adhesion to Caco-2 cells. Finally, PCR-DGGE analysis of the V2-V3 regions of the 16S rRNA gene confirmed the transit of the two selected lactobacilli through the gastrointestinal tract (GIT). Microbiome profiling via the Illumina MiSeq platform revealed the prevalence of Lactobacillus spp. in the gut microbiota of the Lactobacillus-treated rats, even on the 10th day after the Lactobacillus application, compared to the microbiota of the healthy and AlCl3-exposed rats before Lactobacillus treatment. CONCLUSION:The combined application of Lb. plantarum SF9C and Lb. brevis SF9B was able to influence the intestinal microbiota composition in rats, which was reflected in the increased abundance of Lactobacillus genus, but also in the altered abundances of other bacterial genera, either in the model of healthy or aberrant gut microbiota of rats. The antibacterial activity and capacity to withstand in GIT conditions contributed to the functional aspects of SF9C and SF9B strains that could be incorporated in the probiotic-containing functional foods with a possibility to positively modulate the gut microbiota composition.