Mechanism of miR-15a-5p regulation of Wnt pathway in paraquat-induced pulmonary fibrosis
10.3760/cma.j.issn.1671-0282.2024.08.009
- VernacularTitle:miR-15a-5p调控Wnt通路在百草枯致肺纤维化中的机制
- Author:
Jing WANG
1
;
Xiaohang JI
;
Mengmeng WANG
;
Wei ZHANG
;
Weichao DING
;
Juan CHEN
;
Jing FENG
;
Jiankang MENG
;
Zhaorui SUN
;
Shinan NIE
Author Information
1. 南京大学医学院附属金陵医院（中国人民解放军东部战区总医院）急诊医学科，南京 210002
- Keywords:
miRNA;
miR-15a-5p;
Paraquat;
Pulmonary fibrosis;
16HBE cells;
Wnt signaling pathway;
Wnt3α;
β-catenin;
EMT
- From:
Chinese Journal of Emergency Medicine
2024;33(8):1128-1133
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To investigate the effect and molecular mechanism of miR-15a-5p regulation Wnt signaling pathway in PQ-induced pulmonary fibrosis.Methods:The PQ-induced 16HBE cell model was constructed, high-throughput miRNA chip and RT-qPCR were used to screen for miR-15a-5p with significant differences. The experimental groups were as follows: NC group (normal control);no special treatment; PQ group: 50 μmol/L PQ treated cells for 72 h; miR-15a-5p group: 16HBE stable cell lines transfected with miR-15a-5p overexpressing lentivirus; miR-15a-5p+PQ group: Stable cell lines were treated with 50 μmol/L PQ for 72 h. The expression of Wnt pathway-related genes Wnt3α and β-catenin, fibroblast marker genes Collagen I, Vimentin and α SMA, epithelial marker genes Occludin and CK18 were detected by RT-qPCR and Western blot. The mice model of PQ-induced pulmonary fibrosis was constructed, and the protein expression and lung tissue injury were detected by Western blot, HE staining and immunohistochemistry. Data were expressed as mean ± standard deviation, and independent sample t-test was used to analyze the data between the two groups. Results:The expressions of Wnt3α, β-catenin, fibroblast marker genes Collagen I, Vimentin and α SMA significantly up-regulated in cell injury models ( P<0.05), the epithelial cell marker genes Occludin and CK18 significantly down-regulated ( P<0.05), overexpression of miR-15a-5p could inhibit the expression of Wnt3α and alleviated the EMT induced by PQ. In animal models, Wnt3α, β-catenin, fibroblast marker genes Collagen I, Vimentin and α SMA significantly increased ( P<0.01), the structure of lung tissue was disordered and fibrosis occurred, overexpression of miR-15a-5p inhibited the expression of Wnt3α protein ( P<0.05) and ameliorated lung tissue injury. Conclusions:miR-15a-5p ameliorates PQ-induced lung injury by modulating the Wnt3α/β-catenin signaling pathway, thereby inhibiting the development of pulmonary fibrosis.
