Vasodilation of Isolated Vessels and the Isolation of the Extracellular Matrix of Tight-skin Mice
| Autor: | John G. Krolikowski, Kirkwood A. Pritchard, Paul S. Pagel, Omoshalewa Bamkole, Dorothee Weihrauch, Janine A. Struve, Nicole L. Lohr, Tahniyath Zaman, Deron W. Jones |
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| Rok vydání: | 2017 |
| Předmět: |
Pathology
medicine.medical_specialty General Chemical Engineering Apoptosis Inflammation Vasodilation Article General Biochemistry Genetics and Molecular Biology Extracellular matrix Mice Fibrosis medicine Extracellular Animals Myocyte Cell Proliferation General Immunology and Microbiology business.industry General Neuroscience medicine.disease Immunohistochemistry Molecular biology Extracellular Matrix Mice Inbred C57BL Cytosol Models Animal Endothelium Vascular medicine.symptom business |
| Zdroj: | Journal of Visualized Experiments. |
| ISSN: | 1940-087X |
| DOI: | 10.3791/55036 |
| Popis: | The interferon regulatory factor 5 (IRF5) is crucial for cells to determine if they respond in a pro-inflammatory or anti-inflammatory fashion. IRF5's ability to switch cells from one pathway to another is highly attractive as a therapeutic target. We designed a decoy peptide IRF5D with a molecular modeling software for designing small molecules and peptides. IRF5D inhibited IRF5, reduced alterations in extracellular matrix, and improved endothelial vasodilation in the tight-skin mouse (Tsk/+). The Kd of IRF5D for recombinant IRF5 is 3.72 ± 0.74 x 10-6 M as determined by binding experiments using biolayer interferometry experiments. Endothelial cells (EC) proliferation and apoptosis were unchanged using increasing concentrations of IRF5D (0 to 100 µg/mL, 24 h). Tsk/+ mice were treated with IRF5D (1 mg/kg/d subcutaneously, 21 d). IRF5 and ICAM expressions were decreased after IRF5D treatment. Endothelial function was improved as assessed by vasodilation of facialis arteries from Tsk/+ mice treated with IRF5D compared to Tsk/+ mice without IRF5D treatment. As a transcription factor, IRF5 traffics from the cytosol to the nucleus. Translocation was assessed by immunohistochemistry on cardiac myocytes cultured on the different cardiac extracellular matrices. IRF5D treatment of the Tsk/+ mouse resulted in a reduced number of IRF5 positive nuclei in comparison to the animals without IRF5D treatment (50 µg/mL, 24 h). These findings demonstrate the important role that IRF5 plays in inflammation and fibrosis in Tsk/+ mice. |
| Databáze: | OpenAIRE |
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