[Functional polymorphism of NRF2 gene promoter -617C/A in lipopolysaccharide-stimulated peripheral blood mononuclear cellular inflammatory response in patients with alcoholic liver disease].

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).

Conclusion: 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.