The aim of this study was to characterize an in vitro modulating effect of three commensal Lactobacillus strains on cellular differentiation of non-transformed crypt-like rat small intestinal cell line IEC-18. IEC-18 was grown on extracellular matrix, with or without presence of Lactobacillus strains. Gene expression of IEC-18 bacterial detection system - such as Toll-like receptors TLR-2, TLR-4, signal adapter MyD88, cytoplasmic NOD2 receptor, inflammatory cytokines IL-18, IL-1beta, chemokine IL-8 and enzyme caspase-1 - was evaluated using real-time PCR. Expression and localization of TLR-2, TLR-4, IL-18 and caspase-1 proteins was demonstrated by Western blotting and immunofluorescent staining. Secretion of IL-18 to apical and basolateral surfaces was assayed by ELISA. Our results suggested that L. casei LOCK0919 accelerated differentiation of IEC-18 by stimulating TLR-2, TLR-4, MyD88, IL-18, caspase-1 mRNAs and proteins. L. casei LOCK0919 increased expression and transfer of villin and beta-catenin from cytoplasm to cell membrane. Presence of L. rhamnosus LOCK0900 resulted in detachment of IEC-18 layer from extracellular matrix leading to induction of IL-1beta, of TLR-2 and IL-8 mRNAs and stimulation of MyD88, caspase-1 and cytosolic receptor NOD2 mRNAs. L. rhamnosus LOCK0908 was not recognized by TLR-2 or TLR-4 receptors. Lactobacilli-IEC-18 crosstalk enhanced immune and barrier mucosal functions.
- MeSH
- beta-katenin biosyntéza MeSH
- buněčná diferenciace účinky léků MeSH
- cytokiny biosyntéza MeSH
- epitelové buňky účinky léků MeSH
- interleukin-18 biosyntéza MeSH
- kaspasa 1 biosyntéza MeSH
- krysa rodu rattus MeSH
- Lacticaseibacillus rhamnosus * MeSH
- Lactobacillus casei * MeSH
- messenger RNA biosyntéza MeSH
- mikrofilamentové proteiny biosyntéza MeSH
- probiotika farmakologie MeSH
- regulace genové exprese účinky léků MeSH
- střevní sliznice cytologie účinky léků MeSH
- subcelulární frakce metabolismus MeSH
- toll-like receptory biosyntéza účinky léků MeSH
- zvířata MeSH
- Check Tag
- krysa rodu rattus MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
One of the promising approaches in the therapy of ulcerative colitis is administration of butyrate, an energy source for colonocytes, into the lumen of the colon. This study investigates the effect of butyrate producing bacterium Clostridium tyrobutyricum on dextran sodium sulphate (DSS)-induced colitis in mice. Immunocompetent BALB/c and immunodeficient severe combined immunodeficiency (SCID) mice reared in specific-pathogen-free (SPF) conditions were treated intrarectally with C. tyrobutyricum 1 week prior to the induction of DSS colitis and during oral DSS treatment. Administration of DSS without C. tyrobutyricum treatment led to an appearance of clinical symptoms - bleeding, rectal prolapses and colitis-induced increase in the antigen CD11b, a marker of infiltrating inflammatory cells in the lamina propria. The severity of colitis was similar in BALB/c and SCID mice as judged by the histological damage score and colon shortening after 7 days of DSS treatment. Both strains of mice also showed a similar reduction in tight junction (TJ) protein zonula occludens (ZO)-1 expression and of MUC-2 mucin depression. Highly elevated levels of cytokine tumour necrosis factor (TNF)-α in the colon of SCID mice and of interleukin (IL)-18 in BALB/c mice were observed. Intrarectal administration of C. tyrobutyricum prevented appearance of clinical symptoms of DSS-colitis, restored normal MUC-2 production, unaltered expression of TJ protein ZO-1 and decreased levels of TNF-α and IL-18 in the descending colon of SCID and BALB/c mice, respectively. Some of these features can be ascribed to the increased production of butyrate in the lumen of the colon and its role in protection of barrier functions and regulation of IL-18 expression.
- MeSH
- akutní nemoc MeSH
- antigeny CD11b biosyntéza genetika MeSH
- aplikace rektální MeSH
- bakteriální translokace MeSH
- butyráty metabolismus MeSH
- Clostridium tyrobutyricum fyziologie MeSH
- fosfoproteiny biosyntéza genetika MeSH
- imunokompetence MeSH
- interleukin-18 biosyntéza genetika MeSH
- kolon metabolismus mikrobiologie patologie MeSH
- mastné kyseliny metabolismus MeSH
- membránové proteiny biosyntéza genetika MeSH
- mucin 2 biosyntéza genetika MeSH
- muciny biosyntéza MeSH
- myši inbrední BALB C MeSH
- myši SCID MeSH
- myši MeSH
- organismy bez specifických patogenů MeSH
- probiotika terapeutické užití MeSH
- síran dextranu toxicita MeSH
- těžká kombinovaná imunodeficience genetika imunologie MeSH
- TNF-alfa biosyntéza genetika MeSH
- ulcerózní kolitida chemicky indukované genetika imunologie mikrobiologie patologie MeSH
- zvířata MeSH
- Check Tag
- myši MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
It has been well recognized that the promoter polymorphisms of interleukin-18 (IL-18) influence the level of cytokine expression. In our previously published data, we showed constitutive IL-18 expression in the epithelium of renal distal tubules in patients after kidney transplantation and significantly elevated IL-18 expression during acute rejection. In this study, we evaluated the clinical significance of two functional promoter polymorphisms of the IL-18 gene at positions -607 A/C (rs1946518) and -137 C/G (rs187238) in patients after kidney transplantation and looked for associations with the onset of graft function and the incidence of rejection episodes. Promoter polymorphisms in 124 patients and 103 unrelated controls were evaluated by sequence-specific primer polymerase chain reaction and the allele, genotype and haplotype frequencies were statistically correlated. We found a statistically different distribution of the allele frequency of -607 A/C polymorphism between patients with immediate or delayed onset of kidney graft function. Data showed that the C allele, which contributes to higher IL-18 expression, is more frequent in patients with delayed onset of function (P = 0.03, odds ratio = 1.93; 95% confidence interval = 1.15-3.25). A/C single nucleotide polymorphisms of the IL-18 promoter at position -607 may influence the onset of early kidney allograft function.
- MeSH
- časové faktory MeSH
- distální tubuly ledvin metabolismus MeSH
- epitelové buňky metabolismus MeSH
- financování organizované MeSH
- homologní transplantace MeSH
- interleukin-18 biosyntéza genetika MeSH
- jednonukleotidový polymorfismus MeSH
- lidé středního věku MeSH
- lidé MeSH
- obnova funkce genetika MeSH
- přežívání štěpu genetika MeSH
- promotorové oblasti (genetika) genetika MeSH
- regulace genové exprese genetika MeSH
- rejekce štěpu genetika metabolismus MeSH
- retrospektivní studie MeSH
- transplantace ledvin MeSH
- Check Tag
- lidé středního věku MeSH
- lidé MeSH
- mužské pohlaví MeSH
- ženské pohlaví MeSH
- MeSH
- amnion mikrobiologie ultrastruktura MeSH
- finanční podpora výzkumu jako téma MeSH
- fluorescenční protilátková technika MeSH
- imunohistochemie MeSH
- infekce vyvolané Escherichia coli metabolismus MeSH
- interleukin-18 biosyntéza genetika MeSH
- prasata MeSH
- těhotenství MeSH
- transmisní elektronová mikroskopie MeSH
- virulence MeSH
- zvířata MeSH
- Check Tag
- těhotenství MeSH
- ženské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- srovnávací studie MeSH
