Journal
CELLULAR PHYSIOLOGY AND BIOCHEMISTRY
Volume 41, Issue 4, Pages 1596-1604Publisher
KARGER
DOI: 10.1159/000470893
Keywords
miR-136-5p; Astrocyte; IL-17; A20
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Funding
- National Natural Science Foundation of China [81560351]
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Background/Aims: This study focused on investigating the regulatory mechanism of miR-136-5p in mouse astrocytes stimulated with interleukin-17(IL-17). Methods: C57BL/6 mouse astrocytes were stimulated with IL-17 (100ng/m1) for various periods of time (0-48 hours) and at various doses (0-200 ng), and the expression levels of inflammatory cytokine and chemokine genes (IL-6, [NF-alpha, MCP-1, MCP-5 and MIP-2) were then detected by real-time PCR. The expression of the A20 gene was measured with real-time PCR in cells that were stimulated with IL-17 (50 ng/ml) for various periods of time (0-48 hours). C57BL/6 mouse astrocytes were transfected with Ctrl-anti-miR-136-5p or LNA-anti-miR-136-5p for 48 h. Thereafter, the cells were stimulated with or without IL-17 (50ng/m1) for 6 h. The level of A20 protein (TNF alpha-induced protein 3, TNFAIP3) was detected by Western blot analysis. Results: (1) Compared with the DMEM control group, within six hours. IL-17 stimulation significantly increased the expression levels of inflammatory cytokine and chemokine genes and clearly decreased the expression level of the A20 protein. (2) Without IL-17 stimulation, the expression level of the miR-136-5p gene was significantly decreased, whereas in the miR-136-5p-inhibition group, the A20 protein expression was elevated. IL-17 stimulation slightly decreased the expression of the A20 protein in the miR-136-5p-inhibition group, but it was still slightly higher than in the control group. Conclusion: This study demonstrated that miR-136-5p affected the expression of A20 in IL-17-stimulated astrocytes. (C) 2017 The Author(s) Published by S. Karger AG, Basel
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