title: [The effect of transfected microRNA-146a on expression of interleukin-1 receptor-associated kinase 1 and tumor necrosis factor receptor-associated factor 6 in alveolar macrophages].
abstract: OBJECTIVE
  To observe the effect of transfected microRNA-146a (miR-146a) on expression of interleukin-1 receptor-associated kinase 1 (IRAK-1) and tumor necrosis factor receptor-associated factor 6 (TRAF-6) in alveolar macrophages, and to explore the regulatory mechanism of miR-146a in the inflammatory response of alveolar macrophages.
  
  
  METHODS
  Alveolar macrophages NR8383 were cultured and divided into two groups: transfected miR-146a mimic group was transfected 50 nmol/L Pre-miR miR-146a precursors and the negative control group was transfected Cy3-labeled Pre-miR negative control. Cells were collected at 24 hours after transfection. The miR-146a and the mRNA expression of IRAK-1 and TRAF-6 were detected by real-time fluorescent quantitation reverse transcription-polymerase chain reaction (RT-qPCR), and the protein expression of IRAK-1 and TRAF-6 was assayed by Western Blot.
  
  
  RESULTS
  Compared with negative control group, the expression of miR-146a was upregulated by (24.55±6.14) fold compared with miR-146a mimic group (t=-9.353, P=0.001). The mRNA expressions of IRAK-1 and TRAF-6 in miR-146a mimic group were upregulated by (1.16±0.10) fold (t=2.701, P=0.054) and (1.19±0.16) fold (t=2.032, P=0.112) , respectively, compared with that of negative control group, but the protein levels of IRAK-1 and TRAF-6 were decreased by 73.0% (t=-9.353, P=0.001) and 64.1% (t=-6.839, P=0.002), respectively .
  
  
  CONCLUSIONS
  miR-146a mimic was successfully transfected into the alveolar macrophage NR8383. The overexpression of miR-146a in alveolar macrophages can down-regulate the expression of IRAK-1 and TRAF-6 in protein translation levels, and its mechanism may be related with inhibition of protein translation.
authors.affiliation: State Drug Clinical Trial Agency, the First Affiliated Hospital of Nanchang University, Nanchang 330006, Jiangxi, China. Corresponding author: Qian Kejian, Email: qkj0607@sohu.com.
authors.forename: Ping
authors.initials: P
authors.lastname: Huang
authors.forename: Fen
authors.initials: F
authors.lastname: Liu
authors.forename: Zhenguo
authors.initials: Z
authors.lastname: Zeng
authors.forename: Caixue
authors.initials: C
authors.lastname: Huang
authors.forename: Qiang
authors.initials: Q
authors.lastname: Shao
authors.forename: Liang
authors.initials: L
authors.lastname: Xia
authors.forename: Kemin
authors.initials: K
authors.lastname: Jie
authors.forename: Kejian
authors.initials: K
authors.lastname: Qian
mesh_terms: D000818:Animals; D002478:Cells, Cultured; D053592:Interleukin-1 Receptor-Associated Kinases / Q000378:metabolism*; D016676:Macrophages, Alveolar / Q000166:cytology / Q000378:metabolism*; D035683:MicroRNAs / Q000235:genetics*; D051381:Rats; D048029:TNF Receptor-Associated Factor 6 / Q000378:metabolism*; D014162:Transfection
chemical_list: C554180:MIRN146 microRNA, rat; D035683:MicroRNAs; D048029:TNF Receptor-Associated Factor 6; C547807:IRAK1 protein, rat; D053592:Interleukin-1 Receptor-Associated Kinases