A New Hybrid Protein Is a Novel Regulator for Experimental Colitis in Rats
| Autor: | Masaki Ikemoto, Kohki Okada |
|---|---|
| Rok vydání: | 2021 |
| Předmět: |
Male
COVID-19 Lipopolysaccharide Recombinant Fusion Proteins experimental colitis Interleukin-1beta Immunology Flow cytometry chemistry.chemical_compound medicine Animals Calgranulin B Immunology and Allergy Calgranulin A RNA Messenger Rats Wistar hybrid protein NF-κB pathway Gel electrophoresis medicine.diagnostic_test Interleukin-6 Tumor Necrosis Factor-alpha Macrophages Dextran Sulfate Transcription Factor RelA Macrophage Activation Colitis Molecular biology Actins In vitro Rats Blot Real-time polymerase chain reaction chemistry Original Article Tumor necrosis factor alpha Signal transduction |
| Zdroj: | Inflammation |
| ISSN: | 1573-2576 0360-3997 |
| DOI: | 10.1007/s10753-021-01537-5 |
| Popis: | We newly developed a hybrid protein, tentatively named rMIKO-1, using gene technology. We herein investigated the effects of rMIKO-1 on activated macrophages and discussed its potential as a suppressor of experimental colitis. Fluorescent microscopy was used to observe the dynamic mobility of rMIKO-1 in macrophages. Sodium dodecyl sulfate–polyacrylamide gel electrophoresis, Western blotting, fluorescent immunochemical staining, flow cytometry, enzyme-linked immunosorbent assays, a polymerase chain reaction/quantitative polymerase chain reaction, and hematoxylin and eosin staining were conducted to assess the potential activity of rMIKO-1. A large amount of bleeding was observed in rats treated with 5% dextran sulfate sodium (DSS) alone on day 8 after treatment initiation, but not in those treated with 5% DSS plus rMIKO-1. In the in vitro assay, rMIKO-1 rapidly bound to macrophages, immediately entered cells by an unknown mechanism, and then migrated inside the nucleus. This result suggests that rMIKO-1 plays important immunological roles in the nucleus. Despite the activation of macrophages by lipopolysaccharide, the mRNA expression of pro-inflammatory cytokines, such as tumor necrosis factor-α, interleukin-6, and interleukin-1β, was significantly suppressed in macrophages preliminarily treated with rMIKO-1 for 1 h. Complexes of rMIKO-1 with nuclear factor-kappa B (NF-κB)/p65 and β-actin formed in activated macrophages, which attenuated experimental colitis in rats. These results strongly suggest that rMIKO-1 negatively regulates excessively activated macrophages through the NF-κB/p65 signaling pathway. Therefore, rMIKO-1 is a novel suppressor of experimental colitis in rats through the negative regulation of activated macrophages. |
| Databáze: | OpenAIRE |
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