Хирургические аспекты язвенного гастродуоденального кровотечения

ГЕМОРРАГИИ /хирГЕМОСТАЗ ЭНДОСКОПИЧЕСКИЙПЕПТИЧЕСКАЯ ЯЗВА /хи

CYP2J2 is an LPS responsive gene: CYP2J2 products feedback to inhibit TNFα release.

<p>Time course of CYP2J2 mRNA (A) and protein (B) in human peripheral blood mononuclear cells treated with LPS (10 µg/ml). CYP2J2 mRNA was measured by Taqman RT-PCR and expressed as fold of the LPS induced CYP2J2 expression at 4 h. Data shows data points and mean±s.e.m from 4 individual donors. CYP2J2 protein determined by Western blot was compared to that of β-actin. This data is representative of n = 4 separate donors. (C) Effect of 11,12-EET, or 14,15-EET and (D) 9,10-EPOME, or 12,13-EPOME, on basal and LPS (10 µg/ml; 7 h) induced TNFα release from THP-1 monocytes. TNFα release (pg/ml) in the supernatant was measured by ELISA. The data presents the mean±s.e.m. of n = 8 replicates from 3 separate experiments. *denotes p<0.05 control or LPS and EET treatments, and † denotes p<0.05 between control and LPS, by one-way ANOVA and Bonferroni’s post test.</p

CYP2J2 regulates <i>L. monocytogenes</i> phagocytosis.

<p>(A) Epoxygenase inhibition (SKF525A 10 µM; SKF; 24 h) inhibits the infection of <i>L. monocytogenes</i> into THP-1 derived macrophages, expressed as % of bacteria added at 0 h. (B) Right panel shows representative fluorescent micrographs and left panel image analysis of GFP<i>-L. monocytogenes</i> in THP-1 derived macrophages at 7 h. THP-1 derived macrophages were induced by PMA (100 nM; 48 h). 11,12-EET (1 µM) reversed the SKF525A (10 µM) mediated reduction in <i>L. monocytogenes</i> infection. Compounds were given as a 24 h pretreatment before addition of <i>L. monocytogenes</i>. Data represents mean±s.e.m. as a % of control from n = 3–6 separate experiments. * denotes p<0.05 between control and treatments by paired t-test (A) or one-sample t-test (B).</p

Crohn’s disease macrophages do not induce CYP2J2 in response to <i>E. coli</i> stimulation.

<p>(A) unpaired analysis and (B) paired analysis of CYP2J2 mRNA induction in macrophages from Crohn’s disease patients and matched controls in response to heat killed <i>E. coli</i> (2.5×10⁵; 4 h). CYP2J2 mRNA was measured by Taqman RT-PCR and expressed as dCT (4 h). Data shows data points and mean±s.e.m from 7–13 individual donors (A) or 7 paired control and 5 paired Crohn’s disease samples (B). *denotes p<0.05 between control and <i>E. coli</i> treatment, by Mann-Whitney U test.</p

The multicopy sRNA LhrC controls expression of the oligopeptide-binding protein OppA in <i>Listeria monocytogenes</i>

<div><p><i>Listeria monocytogenes</i> is the causative agent of the foodborne disease listeriosis. During infection, <i>L. monocytogenes</i> produces an array of non-coding RNAs, including the multicopy sRNA LhrC. These five, nearly identical sRNAs are highly induced in response to cell envelope stress and target the virulence adhesin <i>lapB</i> at the post-transcriptional level. Here, we demonstrate that LhrC controls expression of additional genes encoding cell envelope-associated proteins with virulence function. Using transcriptomics and proteomics, we identified a set of genes affected by LhrC in response to cell envelope stress. Three targets were significantly down-regulated by LhrC at both the RNA and protein level: <i>lmo2349, tcsA</i> and <i>oppA</i>. All three genes encode membrane-associated proteins: A putative substrate binding protein of an amino acid ABC transporter (Lmo2349); the CD4+ T cell-stimulating antigen TcsA, and the oligopeptide binding protein OppA, of which the latter 2 are required for full virulence of <i>L. monocytogenes</i>. For OppA, we show that LhrC acts by direct base paring to the ribosome binding site of the <i>oppA</i> mRNA, leading to an impediment of its translation and a decreased mRNA level. The sRNA-mRNA interaction depends on 2 of 3 CU-rich regions in LhrC allowing binding of 2 <i>oppA</i> mRNAs to a single LhrC molecule. Finally, we found that LhrC contributes to infection in macrophage-like cells. These findings demonstrate a central role for LhrC in controlling the level of OppA and other virulence-associated cell envelope proteins in response to cell envelope stress.</p></div