The ROS/GRK2/HIF-1 α /NLRP3 Pathway Mediates Pyroptosis of Fibroblast-Like Synoviocytes and the Regulation of Monomer Derivatives of Paeoniflorin.
Zhongyang HongXianzheng ZhangTianjing ZhangLing HuRuijin LiuPan WangHan WangQianqian YuDan MeiZiyang XueFeng ZhangLing-Ling ZhangPublished in: Oxidative medicine and cellular longevity (2022)
Hypoxia is an important factor in the development of synovitis in rheumatoid arthritis (RA). The previous study of the research group found that monomeric derivatives of paeoniflorin (MDP) can alleviate joint inflammation in adjuvant-induced arthritis (AA) rats by inhibiting macrophage pyroptosis. This study revealed increased levels of hypoxia-inducible factor- (HIF-) 1 α and N-terminal p30 fragment of GSDMD (GSDMD-N) in fibroblast-like synoviocytes (FLS) of RA patients and AA rats, while MDP significantly inhibited their expression. Subsequently, FLS were exposed to a hypoxic environment or treated with cobalt ion in vitro. Western blot and immunofluorescence analysis showed increased expression of G protein-coupled receptor kinase 2 (GRK2), HIF-1 α , nucleotide-binding oligomerization segment-like receptor family 3 (NLRP3), ASC, caspase-1, cleaved-caspase-1, and GSDMD-N. Electron microscopy revealed FLS pyroptosis after exposure in hypoxia. Next, corresponding shRNAs were transferred into FLS to knock down hypoxia-inducible factor- (HIF-) 1 α , and in turn, NLRP3 and western blot results confirmed the same. The enhanced level of GSDMD was reversed under hypoxia by inhibiting NLRP3 expression. Knockdown and overexpression of GRK2 in FLS revealed GRK2 to be a positive regulator of HIF-1 α . Levels of GRK2 and HIF-1 α were inhibited by eliminating excess reactive oxygen species (ROS). Furthermore, MDP reduced FLS pyroptosis through targeted inhibition of GRK2 phosphorylation. According to these findings, hypoxia induces FLS pyroptosis through the ROS/GRK2/HIF-1 α /NLRP3 pathway, while MDP regulates this pathway to reduce FLS pyroptosis.
Keyphrases
- nlrp inflammasome
- endothelial cells
- rheumatoid arthritis
- reactive oxygen species
- cell death
- poor prognosis
- high glucose
- binding protein
- disease activity
- dna damage
- signaling pathway
- adipose tissue
- oxidative stress
- single cell
- electron microscopy
- transcription factor
- cell proliferation
- ankylosing spondylitis
- end stage renal disease
- systemic lupus erythematosus
- high resolution
- wound healing
- interstitial lung disease
- idiopathic pulmonary fibrosis
- fluorescent probe
- patient reported outcomes
- diabetic rats
- protein kinase
- single molecule
- simultaneous determination
- molecularly imprinted
- living cells