title: The Effects of miR-136-5p-Mediated Regulation of A20 in Astrocytes from Cultured Spinal Cord Cultured Cells In Vitro.
abstract: OBJECTIVE
  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/ml) 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, TNF-α, 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/ml) for 6 h. The level of A20 protein (TNFα-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.
  
  
  CONCLUSIONS
  This study demonstrated that miR-136-5p affected the expression of A20 in IL-17-stimulated astrocytes.
authors.affiliation: Department of Spine Osteopathia, the First Affiliated Hospital of Guangxi Medical University, Nanning, China.
authors.forename: Xiaoming
authors.initials: X
authors.lastname: Peng
authors.affiliation: Department of Spine Osteopathia, the First Affiliated Hospital of Guangxi Medical University, Nanning, China.
authors.forename: Xiongzhi
authors.initials: X
authors.lastname: Shi
authors.affiliation: Department of Osteopathia, the First Affiliated Hospital of Guangxi Medical University, Nanning, China.|Research Center for Regenerative Medicine, Guangxi Medical University, Nanning, China.
authors.forename: Jinmin
authors.initials: J
authors.lastname: Zhao
authors.affiliation: Department of Spine Osteopathia, the First Affiliated Hospital of Guangxi Medical University, Nanning, China.
authors.forename: Jichen
authors.initials: J
authors.lastname: He
authors.affiliation: Department of Spine Osteopathia, the First Affiliated Hospital of Guangxi Medical University, Nanning, China.
authors.forename: Keke
authors.initials: K
authors.lastname: Li
authors.affiliation: Department of Spine Osteopathia, the First Affiliated Hospital of Guangxi Medical University, Nanning, China.
authors.forename: Zhongxi
authors.initials: Z
authors.lastname: Cen
authors.affiliation: Department of Spine Osteopathia, the First Affiliated Hospital of Guangxi Medical University, Nanning, China.
authors.forename: Yunle
authors.initials: Y
authors.lastname: Wu
authors.affiliation: Department of Spine Osteopathia, the First Affiliated Hospital of Guangxi Medical University, Nanning, China.|Research Center for Regenerative Medicine, Guangxi Medical University, Nanning, China.
authors.forename: Shaohui
authors.initials: S
authors.lastname: Zong
authors.affiliation: College of Public Hygiene of Guangxi Medical University, Nanning, China.|Research Center for Regenerative Medicine, Guangxi Medical University, Nanning, China.
authors.forename: Gaofeng
authors.initials: G
authors.lastname: Zeng
mesh_terms: D000818:Animals; D001253:Astrocytes / Q000166:cytology / Q000378:metabolism*; D002478:Cells, Cultured; D016207:Cytokines / Q000096:biosynthesis / Q000235:genetics; D005786:Gene Expression Regulation; D051379:Mice; D035683:MicroRNAs / Q000235:genetics / Q000378:metabolism*; D013116:Spinal Cord / Q000166:cytology / Q000378:metabolism*; D000072598:Tumor Necrosis Factor alpha-Induced Protein 3 / Q000096:biosynthesis* / Q000235:genetics
chemical_list: D016207:Cytokines; D035683:MicroRNAs; C535074:Mirn136 microRNA, mouse; D000072598:Tumor Necrosis Factor alpha-Induced Protein 3; C513261:Tnfaip3 protein, mouse
keywords: A20; Astrocyte; IL-17; miR-136-5p