title: miR-124 Regulates Amadori-Glycated Albumin-Induced Retinal Microglial Activation and Inflammation by Targeting Rac1.
abstract: To characterize whether the activation of Rac1 is involved in the inflammatory effects produced by Amadori-glycated albumin (AGA) in retinal microglia and to further explore the pathologic pathways of AGA-induced retinal microglial activation and inflammation via a microRNA-dependent mechanism.
  
  
  
  Primary rat retinal microglia were separated and cultured. The levels of TNF-α mRNA and soluble TNF-α produced by the retinal microglia in response to AGA were measured with quantitative RT-PCR (qRT-PCR) and ELISA. In addition, the GTPase activity of Rac1 was measured using a Rac activation assay kit. Luciferase reporter assays were used to validate the regulation of a putative target of microRNA-124 (miR-124).
  
  
  
  Amadori-glycated albumin significantly stimulated the expression of TNF-α mRNA and protein in cultured retinal microglial cells in a dose- and time-dependent manner. MicroRNA-124 expression was consistently suppressed by AGA, and the inhibitory effect was controlled by histone deacetylases (HDACs). Amadori-glycated albumin induced an increase in Rac1 activation in a dose- and time-dependent manner. Furthermore, our data indicated that Rac1 activation-mediated reactive oxygen species production stimulates p65 NF-κB phosphorylation and induces TNF-α release from retinal microglial cells. Finally, we demonstrated that miR-124 directly controls Rac1 expression.
  
  
  
  The current study indicated that AGA-induced retinal microglial activation and inflammation occur via a miR-124-dependent mechanism.
authors.forename: Ning
authors.initials: N
authors.lastname: Dong
authors.forename: Bing
authors.initials: B
authors.lastname: Xu
authors.forename: Hong
authors.initials: H
authors.lastname: Shi
authors.forename: Yan
authors.initials: Y
authors.lastname: Lu
mesh_terms: D000818:Animals; D000831:Animals, Newborn; D015153:Blotting, Western; D002452:Cell Count; D002478:Cells, Cultured; D003921:Diabetes Mellitus, Experimental; D003930:Diabetic Retinopathy / Q000235:genetics* / Q000378:metabolism / Q000473:pathology; D004797:Enzyme-Linked Immunosorbent Assay; D018507:Gene Expression Regulation, Developmental; D017127:Glycation End Products, Advanced; D007249:Inflammation / Q000235:genetics / Q000378:metabolism / Q000473:pathology; D035683:MicroRNAs / Q000096:biosynthesis / Q000235:genetics*; D017628:Microglia / Q000378:metabolism*; D012333:RNA, Messenger / Q000235:genetics; D051381:Rats; D017207:Rats, Sprague-Dawley; D060888:Real-Time Polymerase Chain Reaction; D012165:Retinal Ganglion Cells / Q000378:metabolism* / Q000473:pathology; D012709:Serum Albumin / Q000494:pharmacology*; D015398:Signal Transduction; D014409:Tumor Necrosis Factor-alpha / Q000096:biosynthesis / Q000235:genetics; D020830:rac1 GTP-Binding Protein / Q000096:biosynthesis / Q000235:genetics*; D000093423:Glycated Serum Albumin
chemical_list: D017127:Glycation End Products, Advanced; C548144:MIRN124 microRNA, rat; D035683:MicroRNAs; D012333:RNA, Messenger; D012709:Serum Albumin; D014409:Tumor Necrosis Factor-alpha; C499134:Rac1 protein, rat; D020830:rac1 GTP-Binding Protein; D000093423:Glycated Serum Albumin