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Myostatin/HIF2α-Mediated Ferroptosis is Involved in Skeletal Muscle Dysfunction in Chronic Obstructive Pulmonary
Lijiao Zhang1, Danyang Li1, Chun Chang1
1Department of Respiratory and Critical Care Medicine, Peking University Third Hospital, Beijing, 100191, People's Republic of China.
Objective:
Skeletal muscle dysfunction is an important comorbidity in patients with chronic obstructive pulmonary disease (COPD), and is associated with poor quality of life and reduced survival, but the mechanisms involved remain elusive. Ferroptosis is a newly discovered type of cell death resulting from iron-dependent lipid peroxide accumulation. The purpose of this study was to examine whether ferroptosis is involved in COPD-associated skeletal muscle dysfunction.
Methods:
A mouse model of COPD was established after 24 weeks of cigarette smoke (CS) exposure, and mRNA sequencing, hematoxylin-eosin (H&E) staining, immunostaining (IF), RT-PCR, and Western blot were utilized to identify the changes in gastrocnemius muscles. In vitro, C2C12 myotubes were treated with CS extract (CSE) and evaluated for ferroptosis-related molecules. The pathways regulating ferroptosis were then explored in CSE-stimulated myotubes.
Results:
Compared with controls, COPD mice showed an enriched ferroptosis pathway. Gpx4 was decreased, while hypoxia-inducible factor (Hif) 2α was increased, at gene and protein levels. A reduced level of GSH, but increased cell death, Fe2+, lipid ROS, LPO, and 4-HNE were observed in COPD mice or in CSE-stimulated C2C12 myotubes, which could be ameliorated by ferroptosis inhibitors. The expression of myostatin (MSTN) was enhanced in COPD mice and CSE-stimulated myotubes. MSTN up-regulated HIF2α expression and led to ferroptosis in myotubes, whereas inhibition of MSTN binding to its receptor or inhibition/knockdown of HIF2α resulted in decreased cell death, and partially restored GPX4 and GSH.
Conclusion:
CS exposure induced ferroptosis in vivo and in vitro. Mechanistically, CS-exposure upregulated MSTN which further induced ferroptosis through HIF2α in skeletal muscles, which may contribute to muscle dysfunction through impairing metabolic capacity and decreasing muscle fiber numbers, revealing a potential novel therapeutic target for COPD-related skeletal muscle dysfunction.
Insights
Cigarette smoke exposure triggers ferroptosis, a cell death pathway, in skeletal muscles of chronic obstructive pulmonary disease (COPD) models. This process, driven by myostatin and HIF2α, contributes to muscle dysfunction and offers a potential therapeutic target.
Area of Science:
- Biomedical Science
- Cell Biology
- Respiratory Medicine
Background:
- Skeletal muscle dysfunction is a significant comorbidity in chronic obstructive pulmonary disease (COPD), impacting patient quality of life and survival.
- The precise mechanisms underlying COPD-associated skeletal muscle dysfunction remain largely unknown.
- Ferroptosis, a novel form of programmed cell death driven by iron-dependent lipid peroxidation, is emerging as a key cellular process.
Purpose of the Study:
- To investigate the potential involvement of ferroptosis in the skeletal muscle dysfunction observed in chronic obstructive pulmonary disease (COPD).
- To elucidate the molecular mechanisms linking cigarette smoke exposure to ferroptosis in skeletal muscle.
Main Methods:
- Establishment of a mouse model of COPD through chronic cigarette smoke (CS) exposure.
- Analysis of gastrocnemius muscle changes using mRNA sequencing, hematoxylin-eosin staining, immunostaining, RT-PCR, and Western blot.
- In vitro studies using C2C12 myotubes treated with cigarette smoke extract (CSE) to evaluate ferroptosis markers and regulatory pathways.
Main Results:
- COPD mice exhibited an activated ferroptosis pathway, characterized by decreased GPX4 and increased HIF2α at both gene and protein levels.
- Increased markers of cell death, iron, lipid ROS, and lipid peroxidation were observed in COPD mice and CSE-treated myotubes, which were reversed by ferroptosis inhibitors.
- Myostatin (MSTN) expression was upregulated in COPD models, promoting ferroptosis via HIF2α; inhibiting MSTN or HIF2α reduced cell death and restored key ferroptosis regulators.
Conclusions:
- Cigarette smoke exposure induces ferroptosis in skeletal muscle both in vivo and in vitro.
- Upregulated myostatin (MSTN) drives ferroptosis through HIF2α activation in skeletal muscle following CS exposure.
- This CS-induced ferroptosis pathway represents a potential therapeutic target for mitigating skeletal muscle dysfunction in COPD patients.
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