methylthiouracil and Inflammation

The screening of bioactive compound libraries can be an effective approach for repositioning FDA-approved drugs or discovering new treatments for human diseases. Inhibition of high mobility group box 1 (HMGB1) and restoration of endothelial integrity are emerging as an attractive therapeutic strategies in the management of severe sepsis or septic shock. Here, we examined the effects of methylthiouracil (MTU), used as antithyroid drug, by monitoring the effects on lipopolysaccharide (LPS)- or cecal ligation and puncture (CLP)-mediated release of HMGB1, and on the modulation of HMGB1-mediated inflammatory responses. The anti-inflammatory activities of MTU were determined by measuring permeability, leukocyte adhesion and migration, and the activation of pro-inflammatory proteins in HMGB1-activated HUVECs and mice. MTU inhibited the release of HMGB1 and downregulated HMGB1-dependent inflammatory responses in human endothelial cells. MTU also inhibited HMGB1-mediated hyperpermeability and leukocyte migration in mice. In addition, treatment with MTU reduced CLP-induced release of HMGB1 and sepsis-related mortality and pulmonary injury. Our results indicate that MTUs could be candidate therapeutic agents for various severe vascular inflammatory diseases via the inhibition of the HMGB1 signaling pathway.

Topics: Animals; Anti-Inflammatory Agents; Cell Movement; Disease Models, Animal; HMGB1 Protein; Human Umbilical Vein Endothelial Cells; Humans; Inflammation; Leukocytes; Lipopolysaccharides; Male; Methylthiouracil; Mice; Mice, Inbred C57BL; Sepsis; Signal Transduction

Drug repositioning is used to discover drug candidates to treat human diseases, through the application of drugs or compounds that are approved for the treatment of other diseases. This method can significantly reduce the time required and cost of discovering new drug candidates for human diseases. Previous studies have reported pro-inflammatory responses of endothelial cells to the release of polyphosphate (PolyP). In this study, we examined the anti-inflammatory responses and mechanisms of methylthiouracil (MTU), which is an antithyroid drug, and its effects on PolyP-induced septic activities in human umbilical vein endothelial cells (HUVECs) and mice. The survival rates, septic biomarker levels, behaviour of human neutrophils and vascular permeability were determined in PolyP-activated HUVECs and mice. MTU suppressed the PolyP-mediated vascular barrier permeability, up-regulation of inflammatory biomarkers, adhesion/migration of leucocytes, and activation and/or production of nuclear factor-κB, tumour necrosis factor-α and interleukin-6. Furthermore, MTU demonstrated protective effects on PolyP-mediated lethal death and the levels of the related septic biomarkers. Therefore, these results indicated the therapeutic potential of MTU on various systemic inflammatory diseases, such as sepsis or septic shock.

Topics: Animals; Blood Vessels; Capillary Permeability; Cell Adhesion; Cell Adhesion Molecules; Cell Movement; Disease Models, Animal; Extracellular Signal-Regulated MAP Kinases; Human Umbilical Vein Endothelial Cells; Inflammation; Interleukin-6; Methylthiouracil; Mice, Inbred C57BL; Neutrophils; NF-kappa B; Polyphosphates; Protective Agents; Tumor Necrosis Factor-alpha

This protocol describes microsphere-based protease assays for use in flow cytometry and high-throughput screening. This platform measures a loss of fluorescence from the surface of a microsphere due to the cleavage of an attached fluorescent protease substrate by a suitable protease enzyme. The assay format can be adapted to any site or protein-specific protease of interest and results can be measured in both real time and as endpoint fluorescence assays on a flow cytometer. Endpoint assays are easily adapted to microplate format for flow cytometry high-throughput analysis and inhibitor screening.

Topics: Animals; Biotinylation; Flow Cytometry; Fluorescence Resonance Energy Transfer; Green Fluorescent Proteins; High-Throughput Screening Assays; Humans; Inflammation; Kinetics; Microspheres; Peptide Hydrolases; Peptides; Reproducibility of Results; Temperature

Topics: Adrenalectomy; Animals; Anti-Inflammatory Agents; Catecholamines; Cortisone; Cromolyn Sodium; Dextrans; Drug Antagonism; Drug Synergism; Glucocorticoids; Glucose; Histamine; Histamine H1 Antagonists; Immunogenetics; Inflammation; Methylthiouracil; Mice; p-Methoxy-N-methylphenethylamine; Passive Cutaneous Anaphylaxis; Pedigree; Serotonin Antagonists; Thyroxine; Urticaria

Topics: Aged; Female; Humans; Inflammation; Male; Methylthiouracil; Neoplasms; Skin; Skin Window Technique; Surgical Wound Dehiscence; Triiodothyronine; Wound Healing

Topics: Humans; Inflammation; Lipopolysaccharides; Methods; Methylthiouracil; Skin; Skin Tests; Surgical Wound Dehiscence; Triiodothyronine

Topics: Histological Techniques; Inflammation; Methylthiouracil; Skin; Thiouracil

Topics: Humans; Inflammation; Methylthiouracil; Penicillins; Thiouracil; Thyroiditis