Exploring the effect of LncRNA DANCR to regulate the Keap1-Nrf2/ARE
pathway on oxidative stress in Rheumatoid Arthritis
Abstract
Introduction:This study focused on investigating the effects of LncRNA
DANCR regulation of Keap1-Nrf2/ARE pathway on inflammation and oxidative
stress in RA. Methods:The levels of LncRNA DANCR/miR-486-3p/ Keap1 in
peripheral blood of 30 RA groups and 30 normal subjects were examined,
and the association of LncRNA DANCR with inflammatory indicators of
rheumatoid arthritis was investigated. We construct overexpression
plasmids and small interfering RNAs of LncRNA DANCR to investigate the
relationship between LncRNA DANCR and FLSs viability and migration in
rheumatoid arthritis, as well as the effects on cellular oxidative
stress factors and Keap1-Nrf2/ARE pathway; molecular biology analysis
was used to predict microRNAs that can bind LncRNA DANCR, and luciferase
verified the binding sites of LncRNA DANCR with Keap1 and miR-486-3p; to
further refine the gene and protein expression results, we used RT- qPCR
and immunoblotting assays . Results: In both groups of PBMCs, the
expression levels of LncRNA DANCR and Keap1 mRNA were higher in the
rheumatoid arthritis group than in the normal control group, and the
opposite was true for miR-486-3p; LncRNA DANCR was positively correlated
with TAOC, IL6, RF, IL17, anti-CCP, MDA, and SOD, but not with ESR,
DAS28, IL11, and SOD, DAS28, IL11, ROS, CRP were negatively correlated;
overexpression of lncRNA DANCR stimulated the Keap1-Nrf2/ARE pathway,
decreased the expression of IL10, SOD, TAOC, and increased the
expression levels of MDA, IL11, IL-17, PD-L1, and silencing of lncRNA
DANCR was the opposite, but knockdown of miR- 486-3p or overexpression
of keap1 reversed the expression of the above-mentioned inflammatory and
oxidative factors. In addition, pcDNA-DANCR clearly showed stronger cell
invasion and migration ability and exacerbated its inflammatory
response, we verified their targeting relationship using dual
luciferase. Conclusion: The low-expressed lncRNA DANCR may regulate the
Keap1-Nrf2/ARE pathway and suppress the inflammatory and oxidative
responses in RA patients.