miR-1224-5p Mediates Mitochondrial Damage to Affect Silica-Induced Pulmonary Fibrosis by Targeting BECN1
Abstract
:1. Introduction
2. Results
2.1. miR-1224-5p Is Increased and Mitophagy is Impaired in Mouse Lung Tissues in a Model of Silica-Induced Pulmonary Fibrosis
2.2. Down-Regulated miR-1224-5p Attenuates Silica-Induced Pulmonary Fibrosis and Restores Mitophagy In Vivo
2.3. miR-1224-5p Suppresses Mitophagy by Targeting BECN1
2.4. BECN1 Facilitates PARK2 Translocation to Mitochondria
2.5. PARK2 Knockdown Suppresses Mitophagy, Activates PDGFRs/PI3K/AKT Signaling Pathway and α-SMA Expression in Fibroblasts
3. Discussion
4. Materials and Methods
4.1. Animals and Grouping
4.2. Fibroblast Culture and Treatment
4.3. Histological Analysis
4.4. Quantitative Real Time PCR (qRT-PCR)
4.5. Western Blot
4.6. Luciferase Assays
4.7. miRNA Inhibitor and siRNA Transfection
4.8. Immunofluorescence Staining
4.9. Co-Immunoprecipitation Assay
4.10. Transmission Electron Microscopy
4.11. Mitochondria Isolation
4.12. Statistical Analysis
Supplementary Materials
Acknowledgments
Author Contributions
Conflicts of Interest
Abbreviations
TGF-β1 | Transforming growth factor beta 1 |
BECN1 | Beclin 1, autophagy-related |
PARK2 | Parkinson disease (autosomal recessive, juvenile) 2 |
PDGFR | Platelet-derived growth factor receptor |
TGF-βRII | Transforming growth factor beta receptor 2 |
TNF-α | Tumor necrosis factor alpha |
LPS | Lipopolysaccharide |
α-SMA | Alpha-smooth muscle actin |
PINK1 | PTEN induced putative kinase 1 |
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Groups | Lesion Severity Grade | Average Severity Grade | Lesion Distribution Grade | Average Distribution Grade | ||||||||||
---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
0 | 1 | 2 | 3 | 4 | 5 | 0 | 1 | 2 | 3 | 4 | 5 | |||
Saline | 6 | 0 | 6 | 0 | ||||||||||
Silica | 1 | 1 | 1 | 3 | 3.5 ± 1.8 ** | 1 | 3 | 1 | 1 | 2.5 ± 1.4 ** | ||||
Silica + anta-NC | 1 | 2 | 1 | 2 | 3.2 ± 1.7 ** | 2 | 1 | 2 | 1 | 2.5 ± 1.5 ** | ||||
Silica + anta-1224-5p | 3 | 2 | 1 | 1.0 ± 1.5 ## | 3 | 2 | 1 | 0.8 ± 1.2 ## |
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Wu, Q.; Xu, T.; Liu, Y.; Li, Y.; Yuan, J.; Yao, W.; Xu, Q.; Yan, W.; Ni, C. miR-1224-5p Mediates Mitochondrial Damage to Affect Silica-Induced Pulmonary Fibrosis by Targeting BECN1. Int. J. Mol. Sci. 2017, 18, 2357. https://doi.org/10.3390/ijms18112357
Wu Q, Xu T, Liu Y, Li Y, Yuan J, Yao W, Xu Q, Yan W, Ni C. miR-1224-5p Mediates Mitochondrial Damage to Affect Silica-Induced Pulmonary Fibrosis by Targeting BECN1. International Journal of Molecular Sciences. 2017; 18(11):2357. https://doi.org/10.3390/ijms18112357
Chicago/Turabian StyleWu, Qiuyun, Tiantian Xu, Yi Liu, Yan Li, Jiali Yuan, Wenxi Yao, Qi Xu, Weiwen Yan, and Chunhui Ni. 2017. "miR-1224-5p Mediates Mitochondrial Damage to Affect Silica-Induced Pulmonary Fibrosis by Targeting BECN1" International Journal of Molecular Sciences 18, no. 11: 2357. https://doi.org/10.3390/ijms18112357