The role of 4-octyl itaconate and related mechanisms in lung fibroblast-to-myofibroblast differentiation
10.3760/cma.j.issn.0254-9026.2024.05.010
- VernacularTitle:4-辛基衣康酸在肺成纤维细胞分化中的作用及机制初步探讨
- Author:
Shizhen LI
1
;
Hui GONG
;
Shengyu TAN
;
Xiangyu ZHANG
Author Information
1. 中南大学湘雅二医院老年医学科，长沙 410011
- Keywords:
Lung fibrosis;
4-octyl itaconate;
Oxidative stress;
Nuclear factor-E2 related factor 2
- From:
Chinese Journal of Geriatrics
2024;43(5):603-608
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To investigate the effect of 4-octyl itaconate(4-OI)on transforming growth factor-β1(TGF-β1)-induced lung fibroblast-to-myofibroblast differentiation and related mechanisms.Methods:TGF-β1 was employed to induce the differentiation of the human embryonic lung fibroblast cell line MRC-5, and the effect of 4-OI on lung fibroblast-to-myofibroblast differentiation was examined.Cytotoxicity of 4-OI on MRC-5 cells was detected by the CCK-8 assay.Western blot was used to detect the protein levels of α-smooth muscle actin(α-SMA), collagen 1α1(COL1A1), fibronectin(FN), phosphorylated and total Smad2/3, and nuclear facor-E2 related factor 2(Nrf2).Real-time fluorescence quantitative PCR was used to detect the mRNA expression of α-SMA, COL1A1 and FN.Reactive oxygen species(ROS)levels were assessed by fluorescence microscopy and flow cytometry.Intracellular glutathione(GSH)concentrations were measured by spectrophotometry.Results:Pretreatment with 4-OI was able to inhibit TGF-β1-induced protein overexpression of α-SMA, COL1A1 and FN( F=122.8, 51.5, 27.2, all P<0.05), and increased mRNA levels( F=29.83, 51.62, 94.82, all P<0.01).In addition, 4-OI inhibited TGF-β1-mediated phosphorylation of Smad2/3 proteins in a dose-dependent manner( F=21.80, 36.69, P<0.01 for both).Pretreatment with 4-OI also reversed increased ROS levels( P<0.01)induced by TGF-β1 and enhanced GSH concentrations via disinhibition of TGF-β1( P<0.05).The inhibitory effect of TGF-β1 on Nrf2 expression was alleviated and Nrf2 nuclear translocation was uplifted by 4-OI pretreatment( P<0.05).After silencing Nrf2, 4-OI was unable to inhibit the increased protein expression of COL1A1 induced by TGF-β1, but was still able to inhibit the increased expression of α-SMA and FN protein induced by TGF-β1( P<0.05). Conclusions:4-OI could inhibit lung fibroblast-to-myofibroblast differentiation partially via Nrf 2 activation.
