In vivo inhibition of miR-155 significantly alters post-stroke inflammatory response
| Autor: | Ernesto Caballero-Garrido, Tamar Lordkipanidze, Juan Carlos Pena-Philippides, Tamara Roitbak |
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| Rok vydání: | 2016 |
| Předmět: |
Male
0301 basic medicine Pathology Time Factors Post-stroke inflammation medicine.medical_treatment Microglia/macrophages Mice 0302 clinical medicine CCL12 Microglia medicine.diagnostic_test General Neuroscience Microfilament Proteins MicroRNA Infarction Middle Cerebral Artery dMCAO CREB-Binding Protein CXCL3 medicine.anatomical_structure Cytokine Neurology Phosphatidylinositol-3 4 5-Trisphosphate 5-Phosphatases Cytokines Encephalitis Signal Transduction medicine.medical_specialty Immunology Biology Immunofluorescence miR-155 03 medical and health sciences Cellular and Molecular Neuroscience Suppressor of Cytokine Signaling 1 Protein Western blot In vivo medicine Animals Research Macrophages Calcium-Binding Proteins Antagomirs Molecular biology Mice Inbred C57BL Disease Models Animal MicroRNAs 030104 developmental biology Gene Expression Regulation 030217 neurology & neurosurgery |
| Zdroj: | Journal of Neuroinflammation |
| ISSN: | 1742-2094 |
| DOI: | 10.1186/s12974-016-0753-x |
| Popis: | Background MicroRNA miR-155 is implicated in modulation of the inflammatory processes in various pathological conditions. In our previous studies, we demonstrated that in vivo inhibition of miR-155 promotes functional recovery after mouse experimental stroke. In the present study, we explored if this beneficial effect is associated with miR-155 inhibition-induced alterations in post-stroke inflammatory response. Methods Intravenous injections of a specific miR-155 inhibitor were initiated at 48 h after mouse distal middle cerebral artery occlusion (dMCAO). Temporal changes in the expression of cytokines and key molecules associated with cytokine signaling were assessed at 7, 14, and 21 days after dMCAO, using mouse cytokine gene and protein arrays and Western blot analyses. Electron and immunofluorescence confocal microscopy techniques were used to evaluate the ultrastructural changes, as well as altered expression of specific phenotypic markers, at different time points after dMCAO. Results In the inhibitor-injected mice (inhibitor group), there was a significant decrease in CCL12 and CXCL3 cytokine expression at 7 days and significantly increased levels of major cytokines IL-10, IL-4, IL-6, MIP-1α, IL-5, and IL-17 at 14 days after dMCAO. These temporal changes correlated with altered expression of miR-155 target proteins SOCS-1, SHIP-1, and C/EBP-β and phosphorylation levels of cytokine signaling regulator STAT-3. Electron microscopy showed decreased number of phagocytically active peri-vascular microglia/macrophages in the inhibitor samples. Immunofluorescence and Western blot of these samples demonstrated that expression of leukocyte/ macrophage marker CD45 and phagocytosis marker CD68 was reduced at 7 days, and in contrast, significantly increased at 14 days after dMCAO, as compared to controls. Conclusions Based on our findings, we propose that in vivo miR-155 inhibition following mouse stroke significantly alters the time course of the expression of major cytokines and inflammation-associated molecules, which could influence inflammation process and tissue repair after experimental cerebral ischemia. Electronic supplementary material The online version of this article (doi:10.1186/s12974-016-0753-x) contains supplementary material, which is available to authorized users. |
| Databáze: | OpenAIRE |
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