IL-33 up-regulates eIF3a expression by activating NF-κB signaling pathway to mediate the proliferation and differentiation of mouse pulmonary myofibroblasts and aggravate pulmonary fibrosis.
- Author:
Yunxing GAO
1
;
Yu FU
2
;
Xiao CHEN
3
;
Zepeng LI
3
;
Xiaowei HE
3
;
Xianwei LI
4
Author Information
1. Departments of Medical Microbiology and Immunology of School of Basic Medicine, Wannan Medical College, Wuhu 241002, China.
2. Department of Clinical Medicine of School of Clinical Medicine, Wannan Medical College, Wuhu 241002, China.
3. Department of Pharmacology of School of Pharmacy, Wannan Medical College, Wuhu 241002, China.
4. Department of Pharmacology of School of Pharmacy, Wannan Medical College, Wuhu 241002, China. *Corresponding author, E-mail: wnmclixianwei69@163.com.
- Publication Type:Journal Article
- MeSH:
Animals;
Mice;
Bleomycin/metabolism*;
Cell Differentiation;
Cell Proliferation;
Dimethyl Sulfoxide/pharmacology*;
Fibroblasts;
Interleukin-33/pharmacology*;
Mice, Inbred C57BL;
Myofibroblasts/metabolism*;
NF-kappa B/metabolism*;
NF-KappaB Inhibitor alpha/metabolism*;
Pulmonary Fibrosis;
Signal Transduction
- From:
Chinese Journal of Cellular and Molecular Immunology
2023;39(8):693-700
- CountryChina
- Language:Chinese
-
Abstract:
Objective To investigate the effects and mechanism of Interleukin-33 (IL-33) mediated proliferation and differentiation of pulmonary myofibroblasts (MFbs) in pulmonary fibrosis (PF). Methods C57BL/6 mice were randomly divided into four groups: a control group, a bleomycin (BLM) group, a BLM combined with IL-33 group and a BLM combined with anti-IL-33 antibody group, 12 mice in each group. The PF model was induced by intratracheal injection of BLM (5000 U/kg). The degrees of fibrosis were examined using HE and Masson staining. ELISA was used to measure the plasma levels of IL-33. Immunohistochemical staining was used to measure the expression of alpha smooth muscle actin (α-SMA) in lung tissue. Primary pulmonary fibroblasts were isolated and cultured from lung tissues of mice. The cells were divided into four groups: a control group, an IL-33 group, an IL-33 combined with dimethyl sulfoxide (DMSO) group and an IL-33 combined with pyrrolidine dithiocarbamate (PDTC) group. The cells were treated with DMSO or PDTC for 1 hour and then with IL-33 for 48 hours. Cell proliferation was measured by 5-ethynyl-2'-deoxyuridine (EdU) assay and cell cycle was measured by flow cytometry. TranswellTM assay was used to analyze cell migration. Real-time quantitative PCR was used to measure the expression of collagen type I (Col1), Col3 and α-SMA mRNA. The protein levels of IL-33, Col1, Col3, α-SMA, eukaryotic initiation factor 3a (eIF3a), phosphorylated IκBα (p-IκBα) (total lysate), p-NF-κB p65(total lysate) and NF-κB p65 (nucleus) were measured by Western blot analysis. Results In vivo, compared with the control group, the expressions of IL-33, p-IκBα (total lysate), p-NF-κB p65 (total lysate), NF-κB p65(nucleus), eIF3a, α-SMA, Col1 and Col3 in the BLM group significantly increased. Compared with the BLM group, the expressions of p-IκBα (total lysate), p-NF-κB p65 (total lysate), NF-κB p65 (nucleus), eIF3a, α-SMA, Col1 and Col3 in the IL-33 group increased further and the PF was further aggravated. But the effect of anti-IL-33 antibody was just opposite to that of IL-33. In vitro, IL-33 markedly induced the proliferation and migration of pulmonary fibroblasts, and significantly up-regulated the expression of p-IκBα (total lysate), p-NF-κB p65(total lysate), NF-κB p65 (nucleus), eIF3a, α-SMA, Col1 and Col3. But all these effects of IL-33 were reversed by pyrrolidine dithiocarbamate. Conclusion The results suggest that IL-33 may promote the expression of eIF3a by activating NF-κB signaling pathway, thus inducing the proliferation and differentiation of MFbs and promoting the occurrence and development of PF.
