title: Decreased miR-146a expression in acute ischemic stroke directly targets the Fbxl10 mRNA and is involved in modulating apoptosis.
abstract: BACKGROUND
  miR-146a, a strong pro-apoptotic factor in some pathophysiological processes, is reported to be involved in ischemic stroke (IS), though its role remains unclear. Fbxl10 is an active anti-apoptotic factor and a predicted target of miR-146a. We hypothesized that dysregulation of miR-146a contributes to ischemic injury by targeting Fbxl10.
  
  
  METHODS
  Circulating miRNAs were detected by miRNA microarray and qRT-PCR. miR-146a targets were predicted using bioinformatics and confirmed with a dual luciferase reporter assay. We used an in vitro ischemic model of oxygen-glucose deprivation and reperfusion (OGD/R) to mimic cerebral ischemia/reperfusion (I/R) conditions. Expression of miR-146a, Fbxl10 and Bcl2l2 mRNAs, and Fbxl10 and Bcl2l2 proteins was verified by qRT-PCR and Western blotting. The effects of miR-146a on neuronal cell apoptosis were evaluated by flow cytometry.
  
  
  RESULTS
  A significant reduction in miR-146a expression was observed in acute ischemic stroke (AIS). A dual-luciferase reporter assay showed that Fbxl10, but not Bcl2l2, is a target of miR-146a. Transfection with miR-146a mimics promoted apoptosis in SK-N-SH cells and significantly reduced expression of Fbxl10. Conversely, miR-146a inhibition attenuated OGD/R-induced neuronal cell death and significantly up-regulated Fbxl10 expression.
  
  
  CONCLUSIONS
  miR-146a expression was significantly down-regulated in AIS, and Fbxl10 was identified as a target of miR-146a. Moreover, up-regulation of Fbxl10, a miR-146a target, likely protects neurons from ischemic death.
authors.affiliation: Department of Neurology, The First Affiliated Hospital of Guangxi Medical University in Nanning, China. Electronic address: shenghuali2013@163.com.
authors.forename: Sheng-Hua
authors.initials: SH
authors.lastname: Li
authors.affiliation: Department of Internal Medicine, The First Affiliated Hospital of Guangxi Medical University in Nanning, China.
authors.forename: Lan
authors.initials: L
authors.lastname: Chen
authors.affiliation: Department of Neurology, The First Affiliated Hospital of Guangxi Medical University in Nanning, China.
authors.forename: Xiao-Min
authors.initials: XM
authors.lastname: Pang
authors.affiliation: Department of Neurology, The First Affiliated Hospital of Guangxi Medical University in Nanning, China.
authors.forename: Sheng-You
authors.initials: SY
authors.lastname: Su
authors.affiliation: Department of Neurology, The First Affiliated Hospital of Guangxi Medical University in Nanning, China.
authors.forename: Xia
authors.initials: X
authors.lastname: Zhou
authors.affiliation: Department of Neurology, The First Affiliated Hospital of Guangxi Medical University in Nanning, China.
authors.forename: Chun-Yong
authors.initials: CY
authors.lastname: Chen
authors.affiliation: Department of Neurology, The First Affiliated Hospital of Guangxi Medical University in Nanning, China.
authors.forename: Li-Gang
authors.initials: LG
authors.lastname: Huang
authors.affiliation: Department of Neurology, The First Affiliated Hospital of Guangxi Medical University in Nanning, China.
authors.forename: Jing-Pin
authors.initials: JP
authors.lastname: Li
authors.affiliation: Department of Neurology, The First Affiliated Hospital of Guangxi Medical University in Nanning, China. Electronic address: lilicomet@163.com.
authors.forename: Jing-Li
authors.initials: JL
authors.lastname: Liu
mesh_terms: D000368:Aged; D017209:Apoptosis / Q000502:physiology*; D002545:Brain Ischemia / Q000097:blood* / Q000000981:diagnostic imaging; D045744:Cell Line, Tumor; D044783:F-Box Proteins / Q000097:blood* / Q000235:genetics; D005260:Female; D015870:Gene Expression; D057809:HEK293 Cells; D006801:Humans; D056484:Jumonji Domain-Containing Histone Demethylases / Q000097:blood* / Q000235:genetics; D008297:Male; D035683:MicroRNAs / Q000097:blood* / Q000235:genetics; D008875:Middle Aged; D040081:Protein Array Analysis / Q000379:methods; D012333:RNA, Messenger / Q000097:blood / Q000235:genetics; D020521:Stroke / Q000097:blood* / Q000000981:diagnostic imaging
chemical_list: D044783:F-Box Proteins; C542542:MIRN146 microRNA, human; D035683:MicroRNAs; D012333:RNA, Messenger; D056484:Jumonji Domain-Containing Histone Demethylases; C534524:KDM2A protein, human
keywords: AIS; Fbxl10; OGD/R; miR-146a