title: [Functional polymorphism of NRF2 gene promoter -617C/A in lipopolysaccharide-stimulated peripheral blood mononuclear cellular inflammatory response in patients with alcoholic liver disease].
abstract: OBJECTIVE
  To investigate the influence of NRF2 gene polymorphism at locus -617 on inflammatory response of lipopolysaccharide (LPS)-stimulated peripheral blood mononuclear cells (PBMCs) in patients with alcoholic liver disease (ALD).
  
  
  METHODS
  Venous blood samples from 82 patients with ALD were collected and PBMCs were separated using Ficoll density gradient centrifugation. T cell subgroup was detected by flow cytometry. The polymorphisms in NRF2 gene promoter -617C/A was determined by gene sequencing. According to the results of gene sequencing, patients were divided into non-mutation group (genotype CA and AA) and mutation group (genotype CC). After stimulation with LPS, the expression levels of NRF2, tumor necrosis factor (TNF)α, interleukin (IL)-1β and IL-10 were measured by reverse transcription-PCR (RT-PCR) and enzyme linked immunosorbent assay (ELISA), respectively.
  
  
  RESULTS
  Among the 82 patients with ALD, 32 were homozygous for the C allele (CC), 44 heterozygous (CA), and 6 AA. The frequencies of allele C and A were 65.9% and 34.1%, respectively. There were no differences in clinical data, such as liver function and distribution of T cell subsets between the two groups (all P values >0.05) .Under LPS stimulation, the NRF2 mRNA expression in the non-mutation group was significantly higher than that in the mutation group (P < 0.05). The TNFα, IL-1β mRNA and protein expression in the mutation group were significantly higher than those in the non-mutation group (P < 0.05) and IL-10 mRNA and protein expression of the mutation group was higher than that in the non-mutation group without statistical significance (P > 0.05).
  
  
  CONCLUSIONS
  The gene promoter NRF2-617C mutated to A in LPS-stimulated PBMC of patients with ALD significantly decreases the expression of NRF2 and releases early proinflammatory cytokines.
authors.affiliation: Emergency Medical Department, the First Affiliated Hospital, Wenzhou Medical College, Wenzhou 325000, China.
authors.forename: Guang-ju
authors.initials: GJ
authors.lastname: Zhao
authors.forename: Xiao
authors.initials: X
authors.lastname: Chen
authors.forename: Xiao-lin
authors.initials: XL
authors.lastname: Li
authors.forename: Yi-juan
authors.initials: YJ
authors.lastname: Yu
authors.forename: Guang-liang
authors.initials: GL
authors.lastname: Hong
authors.forename: Qiao-meng
authors.initials: QM
authors.lastname: Qiu
authors.forename: Meng-fang
authors.initials: MF
authors.lastname: Li
authors.forename: Zhong-qiu
authors.initials: ZQ
authors.lastname: Lu
mesh_terms: D000328:Adult; D000368:Aged; D002478:Cells, Cultured; D005260:Female; D005838:Genotype; D006801:Humans; D007249:Inflammation / Q000139:chemically induced / Q000378:metabolism; D007963:Leukocytes, Mononuclear / Q000378:metabolism*; D008108:Liver Diseases, Alcoholic / Q000235:genetics* / Q000378:metabolism*; D008297:Male; D008875:Middle Aged; D051267:NF-E2-Related Factor 2 / Q000235:genetics*; D020641:Polymorphism, Single Nucleotide; D011401:Promoter Regions, Genetic; D012333:RNA, Messenger / Q000235:genetics
chemical_list: D051267:NF-E2-Related Factor 2; C495635:NFE2L2 protein, human; D012333:RNA, Messenger