慢性阻塞性肺疾病大鼠气道成纤维细胞过表达HSG基因对Wnt和MAPK信号通路的影响

doi:10.3969/j.issn.1006-5725.2023.22.011

Abstract

Abstract:

Objective To study the effect of overexpression of HSG gene （MFN2） on Wnt signaling pathway in airway fibroblasts of rats with chronic obstructive pulmonary disease （COPD）. Methods The rat COPD model was induced by protease， and then the airway fibroblasts of COPD model rats were isolated in primary culture. The expression vector of HSG gene was constructed and transfected into rat airway fibroblasts to obtain rat airway fibroblasts overexpressing HSG. In Experiment 1， cells were treated with phorbol ester （TPA）， a Wnt signaling pathway agonist， and the expressions of HSG and Wnt5a were detected by WB and qPCR； In Experiment 2， the cells were treated with EGF factor， an agonist of MAPK signal pathway， and the expressions of HSG and ERK1/2 were detected by WB and qPCR； Moreover， ELISA was used to detect TGF in cell culture medium in each experiment-β1. The contents of PDGF and MMP-9， CCK8， cell proliferation and apoptosis were detected by flow cytometry. Results Airway fibroblasts were successfully isolated from COPD model rats. In Experiment 1 and Experiment 2， WB and qPCR showed that overexpression of HSG could inhibit the expression of genes Wnt5a and ERK1/2. After treatment with corresponding agonists， the expression of these genes was up-regulated and the expression of HSG was inhibited； The results of ELISA showed that overexpression of HSG could reduce TGF-β1. The contents of PDGF and MMP-9， TGF after treatment with corresponding agonists-β1. The contents of PDGF and MMP-9 increased， but overexpression of HSG still had inhibitory effect. CCK8 detection showed that overexpression of HSG could inhibit cell proliferation， while flow cytometry showed that overexpression of HSG could significantly increase the apoptosis rate， and the corresponding agonist treatment would reduce the effect of overexpression of HSG on cell apoptosis. Flow cytometry showed that the effect of overexpression of HSG on cell cycle was not obvious. Conclusion Overexpression of HSG gene in airway fibroblasts of COPD rats can inhibit Wnt， MAPK and other signal pathways， and overexpression of HSG gene can promote fibroblast apoptosis and inhibit cell proliferation. It can be speculated that promoting HSG expression can inhibit airway fibrosis of COPD rats.

Key words: chronic obstructive pulmoriary disease, Wnt signaling pathway, fibroblast, HSG, MAPK signaling pathway

CLC Number:

R563.9

Yi YANG,Xun ZHOU,Jun ZHANG,Bo LI,Xinxing WANG,Xian LUO,Zhengxing. GE. Effects of overexpression of HSG gene on Wnt signaling pathway and MAPK signaling pathway in COPD rat airway fibroblasts[J]. The Journal of Practical Medicine, 2023, 39(22): 2921-2927.

Figures/Tables 6

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Effects of overexpression of HSG gene on Wnt signaling pathway and MAPK signaling pathway in COPD rat airway fibroblasts

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