Study on the Mechanism of miR-93 Targeting MFN2 to Regulate Mitochondrial-associated Endoplasmic Reticulum Membrane Homeostasis and Affect ARDS-induced Pulmonary Fibrosis
10.3870/j.issn.1672-0741.25.04.015
- VernacularTitle:miR-93靶向MFN2调节线粒体相关内质网膜动态平衡影响ARDS肺纤维化的机制研究
- Author:
Ning AN
1
;
Meixia XU
;
Xiaoxia ZHANG
Author Information
1. 华中科技大学同济医学院附属协和医院麻醉与危重病研究所,武汉 430022
- Publication Type:Journal Article
- Keywords:
ARDS pulmonary fibrosis;
miR-93;
MFN2;
endoplasmic reticulum stress
- From:
Acta Medicinae Universitatis Scientiae et Technologiae Huazhong
2025;54(4):491-497
- CountryChina
- Language:Chinese
-
Abstract:
Objective To investigate the regulatory role of miR-93/MFN2 axis in pulmonary fibrosis associated with acute respiratory distress syndrome(ARDS)through endoplasmic reticulum stress(ERS)and mitochondrial function.Methods An in v itro model of ARDS-induced pulmonary fibrosis was established by treating human embryonic lung fibroblasts(HFL-1)with li-popolysaccharide(LPS).Four experimental groups were employed in functional validation:negative control inhibitor(inhibitor NC),miR-93 inhibitor,miR-93 inhibitor+scrambled control siRNA(si-NC),and miR-93 inhibitor+MFN2-targeting siRNA(si-MFN2).Collagen Ⅰ(COLⅠ)secretion was assesed by ELISA and Western blotting;MFN2 and miR-93 expressions were detec-ted by qRT-PCR;Cell proliferation was assessed by CCK-8 assay;Apoptosis was evaluated by flow cytometry;ERS marker(Bip/GRP78,IRE1)levels and mitophagy protein Beclin-1 were detecetd by Western blotting.The miR-93-MFN2 interaction was confirmed via dual-luciferase assays.Results Compared with control group,COL Ⅰ secretion in HFL-1 cells was signifi-cantly increased in LPS group(P＜0.01 vs.control),validating the in vitro pulmonary fibrosis model.Compared to the inhibitor NC group,the miR-93 inhibitor group exhibited significantly upregulated MFN2 expression,decreased HFL-1 cell proliferation,increased apoptosis,and reduced COLⅠ secretion(all P＜0.01).MFN2 was confirmed as a direct target of miR-93 by dual-lu-ciferase assays.Compared with inhibitor NC group,levels of ERS markers(Bip/GRP78,IRE1)were downregulated,and the mi-tophagy marker Beclin-1 was upregulated in miR-93 inhibitor group(P＜0.01).These effects were rescued after co-transfection with miR-93 inhibitor and si-MFN2.Compared to the miR-93 inhibitor group alone,the miR-93 inhibitor+si-MFN2 group showed attenuated suppression of cell proliferation,increased COLⅠ secretion,upregulated ERS markers(Bip/GRP78,IRE1),and downregulated Beclin-1 expression.Conclusion miR-93 exacerbates ARDS-associated pulmonary fibrosis by directly targe-ting and suppressing MFN2.This disruption likely compromised mitochondria-associated ER membrane(MAM)homeosta-sis.Targeting themiR-93/MFN2 axis alleviates fibrosis by attenuating ERS and promoting mitophagy,highlighting its potential as a therapeutic strategy.
