title: Advanced glycation end products activate the miRNA/RhoA/ROCK2 pathway in endothelial cells.
abstract: OBJECTIVE
  AGEs induce endothelial cell dysfunction in HUVECs, resulting in ROS production and triggering apoptosis. This study sought to identify miRNAs involved in AGE-induced endothelial cell injury.
  
  
  METHODS
  Microarray analysis to identify miRNAs altered with AGE stimulation was undertaken, and results were confirmed using real-time quantitative polymerase chain reaction. The interaction of miRNAs with the RhoA and ROCK2 genes was confirmed using luciferase assays, and their effects on expression were determined using Western blot analysis. The effects of AGEs and miRNAs on endothelial cell permeability were assessed.
  
  
  RESULTS
  AGEs induced ROS production and apoptosis of HUVECs (p < 0.05). AGE-induced miR-200b and miR-200c downregulation led to increased expression of their target genes, RhoA and ROCK, respectively. AGE-induced endothelial cell permeability and F-actin expression were significantly reduced with both miR-200b and miR-200c mimics (p < 0.05). Furthermore, AGE-induced stress fiber formation was reduced in cells treated with miR-200b mimics.
  
  
  CONCLUSIONS
  miR-200b and miR-200c are suppressed in AGE-induced endothelial cell injury, resulting in unregulated RhoA/ROCK2 signaling. Further studies are necessary to evaluate the therapeutic value of targeting miRNAs or their target genes for treatment of vascular diseases.
authors.affiliation: Department of Anesthesiology, Zhujiang Hospital, Southern Medical University, Guangzhou, China.
authors.forename: Xiao-Dan
authors.initials: XD
authors.lastname: Wu
authors.forename: Wei-Lin
authors.initials: WL
authors.lastname: Liu
authors.forename: Kai
authors.initials: K
authors.lastname: Zeng
authors.forename: Hong-Yi
authors.initials: HY
authors.lastname: Lei
authors.forename: Qing-Guo
authors.initials: QG
authors.lastname: Zhang
authors.forename: Shu-Qin
authors.initials: SQ
authors.lastname: Zhou
authors.forename: Shi-Yuan
authors.initials: SY
authors.lastname: Xu
mesh_terms: D017209:Apoptosis / Q000187:drug effects; D002199:Capillary Permeability; D017127:Glycation End Products, Advanced / Q000378:metabolism* / Q000494:pharmacology; D061307:Human Umbilical Vein Endothelial Cells / Q000378:metabolism* / Q000473:pathology; D006801:Humans; D035683:MicroRNAs / Q000378:metabolism*; D054460:rho-Associated Kinases / Q000378:metabolism*; D020742:rhoA GTP-Binding Protein / Q000378:metabolism*
chemical_list: D017127:Glycation End Products, Advanced; C533896:MIRN200 microRNA, human; D035683:MicroRNAs; C066737:RHOA protein, human; C515852:ROCK2 protein, human; D054460:rho-Associated Kinases; D020742:rhoA GTP-Binding Protein
keywords: F-actin; RhoA-ROCK; advanced glycation end products; monolayer permeability; reactive oxygen species