title: [Effects of NF-E2-related factor-2 promoter polymorphism on lipopolysaccharide-induced inflammatory responses in macrophages].
abstract: OBJECTIVE
  To explore the effects of NF-E2-related factor-2 (NRF2)-617C/A promoter polymorphism on NRF2 expression as well as lipopolysaccharide-induced inflammatory responses in macrophages.
  
  
  METHODS
  NRF2-617C/A promoter fragments were synthesized by chemical method and cloned into a pUC57 vector. The dul-luciferase reporter assay was employed to determine the activity of promoters. Then recombinant adenoviral vectors were constructed and transfected into macrophages. The expression of Nrf2 was examined by Western blotting and reverse transcription (RT)-PCR. The expressions of tumor necrosis factor-alpha (TNF-α), interleukin-6 (IL-6) and interleukin-10 (IL-10) in macrophages after the stimulation of LPS were determined by enzyme-linked immunosorbent assay (ELISA).
  
  
  RESULTS
  The activity of NRF2-617C promoter-luciferase reporter (FLuc/RLuc activity ratio) was significantly higher than that of NRF2-617A group (0.584 ± 0.016 vs 0.258 ± 0.018, P < 0.05).The NRF2 protein and mRNA levels in -617C group were much higher than those of 617A group (1.123 ± 0.080 vs 0.951 ± 0.057,1.889 ± 0.031 vs 1.647 ± 0.323, both P < 0.05). After the stimulation of LPS, the NRF2 protein expression in macrophages significantly increased (0.584 ± 0.016 vs 0.258 ± 0.018, P < 0.05). Compared with -617A group, there was a significantly higher expression of NRF2 in -617C group (0.671 ± 0.033 vs 0.751 ± 0.014, P < 0.05). Additionally, the productions of IL-6 and IL-10 in -617C group were markedly lower than those in -617A group as well as IL-6/IL-10 (both P < 0.05). However, no significant difference existed in the levels of TNF-α between -617C and -617A groups (P > 0.05).
  
  
  CONCLUSIONS
  The -617C/A promoter polymorphism of NRF2 may influence the NRF2 expression. And it appears to be associated with the LPS-induced inflammatory responses in macrophages.
authors.affiliation: Department of Emergency Medicine, Wenzhou Medical College, Wenzhou, China.
authors.forename: Qiao-meng
authors.initials: QM
authors.lastname: Qiu
authors.forename: Jin-tao
authors.initials: JT
authors.lastname: Zheng
authors.forename: Chao
authors.initials: C
authors.lastname: Nan
authors.forename: Yuan-yuan
authors.initials: YY
authors.lastname: Zhao
authors.forename: Guang-ju
authors.initials: GJ
authors.lastname: Zhao
authors.forename: Guang-liang
authors.initials: GL
authors.lastname: Hong
authors.forename: Huan
authors.initials: H
authors.lastname: Liang
authors.forename: Meng-Fang
authors.initials: MF
authors.lastname: Li
authors.forename: Zhong-Qiu
authors.initials: ZQ
authors.lastname: Lu
mesh_terms: D002478:Cells, Cultured; D006801:Humans; D007249:Inflammation; D016753:Interleukin-10 / Q000378:metabolism; D015850:Interleukin-6 / Q000378:metabolism; D008070:Lipopolysaccharides; D008264:Macrophages / Q000378:metabolism / Q000473:pathology*; D051267:NF-E2-Related Factor 2 / Q000235:genetics*; D020641:Polymorphism, Single Nucleotide; D011401:Promoter Regions, Genetic; D014409:Tumor Necrosis Factor-alpha / Q000378:metabolism
chemical_list: C508609:IL10 protein, human; C508600:IL6 protein, human; D015850:Interleukin-6; D008070:Lipopolysaccharides; D051267:NF-E2-Related Factor 2; C495635:NFE2L2 protein, human; D014409:Tumor Necrosis Factor-alpha; D016753:Interleukin-10