TTP deficiency blocks the anti-inflammatory function of CO in DSS-induced colitis.

<p>After treatment with 2% (w/v) DSS for 7 days, WT and TTP KO mice were exposed to CO at a concentration of 250 ppm for 5 days and analyzed for colitis. A. Clinical scores were assessed as described in Materials and Methods. Data are mean ± SD for 5 mice (**, p<0.01; ***, p<0.005). ns, not significant. B and C. Colon length. B. Representative images of 5 tests conducted in each group. C. Data are mean ± SD for 5 mice (*, p<0.05; **, p<0.01). D. Representative H&E sections of each group from colons of WT and TTP KO mice. E. Neutrophil infiltration into the colon, quantified by measuring MPO activity. Data are mean ± SD for 5 mice (*, p<0.05; **, p<0.01). F. Colonic cytokine/chemokine mRNA levels, analyzed by semi-quantitative RT-PCR. Data represent 1 of 3 independent experiments with similar results. The band densities in the agarose gel were quantified by PhosphorImager, normalized to the internal control GAPDH and expressed as percentage (%) of the value of untreated control cells. Data shown are mean ± SD (n = 3). **, p<0.01; ***, p<0.005. ns, not significant.</p

CORM-2 induces TTP promoter activity and <i>TTP</i> mRNA in macrophages.

<p>A. RAW264.7 cells were incubated with different concentration of CORM-2 or iCORM-2 for 4 h. <i>TTP</i> mRNA expression was determined by semi-quantitative RT-PCR. B. RAW264.7 cells were transfected with pGL3/mTTP P-1309 containing the mouse <i>TTP</i> promoter. After 24 h, cells were treated with different concentrations of CORM-2 for 4 h, and luciferase activity was determined. The level of firefly luciferase activity was normalized using <i>Renilla</i> luciferase activity. The relative luciferase activity is presented as a fold increase over untreated cells. Values are mean ± SD (n = 3). *, <i>p</i><0.05; **, <i>p<</i>0.01.</p

TTP deficiency blocks the anti-inflammatory function of CORM-2 in macrophages.

<p>Peritoneal macrophages were harvested from WT and TTP KO mice. Cells were treated with 1 µg/ml LPS in the presence of 10 µM CORM-2 for 4 h. A. TNF-α in the supernatant of cells was determined by ELISA. Data shown are mean ± SD (n = 3). **, <i>p<</i>0.01. B. The expression levels of <i>TTP</i> and <i>TNF-α</i> mRNA were determined by semi-quantitative RT-PCR. C. Expression of <i>TNF</i> mRNA in macrophages was determined by quantitative real-time PCR at indicated times after the addition of 5 µg/ml actinomycin D. Values are mean ± SD (n = 3). ***, <i>p<</i>0.001. The band densities in the agarose gel were quantified by PhosphorImager, normalized to the internal control GAPDH and expressed as percentage (%) of the value of untreated control cells. Data shown are mean ± SD (n = 3). ***, p<0.005. ns, not significant.</p