The Sphingosine Kinase 1 Inhibitor, PF543, Mitigates Pulmonary Fibrosis by Reducing Lung Epithelial Cell mtDNA Damage and Recruitment of Fibrogenic Monocytes
Abstract
:1. Introduction
2. Results
2.1. PF543 Administered Following Exposure to Either Bleomycin or Asbestos Mitigates Pulmonary Fibrosis
2.2. PF543 Attenuates Asbestos-Induced S1P Production and mtDNA Damage in Mouse Lungs and MLE Cells
2.3. PF543 Diminishes Recruitment of Pro-Fibrotic Mo-AMs
3. Discussion
4. Materials and Methods
4.1. Reagents
4.2. Animals
4.3. Bleomycin Instillation into Mice
4.4. Asbestos Preparation and Instillation into Mice
4.5. Lung Harvest and Histology
4.6. Lung Collagen Detection
4.7. Fibrosis Scoring System
4.8. Bronchoalveolar Lavage (BAL) Cell/Protein Isolation
4.9. PF543 Treatment of Mice
4.10. S1P Determination by Mass Spectrometry
4.11. Quantitative mtDNA Damage Assay via PCR
4.12. Western Blotting
4.13. Cell Culture
4.14. Induction of Transgene Expression in zsGreen Mice
4.15. Cell Isolation, Staining, Flow for Mo-AMs
4.16. Statistical Analysis
Supplementary Materials
Author Contributions
Funding
Conflicts of Interest
Abbreviations
ATF3 | Activating transcription factor 3 |
α-SMA | alpha-smooth muscle actin |
AEC | alveolar epithelial cell |
AT2 | alveolar type II cell |
BER | base excision repair |
Col1A2 | collagen-1A2 |
ER | endoplasmic reticulum |
H&E | hematoxylin and eosin |
HLF | human lung fibroblasts |
mtOgg1tg | human mitochondria-targeted OGG1 subunit 1-alpha transgene |
IPF | idiopathic pulmonary fibrosis |
IP | intra-peritoneal |
IT | intra-tracheal |
MCAT | mitochondrial catalase enforced-expression |
mtDNA | Mitochondrial DNA |
mtOGG1 | mitochondrial 8-oxoguanine-DNA glycosylase 1 |
OGG1 | 8-oxyguanine DNA glycosylase |
PINK1 | PTEN-induced putative kinase 1 |
ROS | Reactive oxygen species |
SPHK1 | sphingosine-kinase 1 |
S1P | sphingosine-1-phosphate |
S1PL | S1P lyase |
TGFβ | transforming growth factor-beta |
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Cheresh, P.; Kim, S.-J.; Huang, L.S.; Watanabe, S.; Joshi, N.; Williams, K.J.N.; Chi, M.; Lu, Z.; Harijith, A.; Yeldandi, A.; et al. The Sphingosine Kinase 1 Inhibitor, PF543, Mitigates Pulmonary Fibrosis by Reducing Lung Epithelial Cell mtDNA Damage and Recruitment of Fibrogenic Monocytes. Int. J. Mol. Sci. 2020, 21, 5595. https://doi.org/10.3390/ijms21165595
Cheresh P, Kim S-J, Huang LS, Watanabe S, Joshi N, Williams KJN, Chi M, Lu Z, Harijith A, Yeldandi A, et al. The Sphingosine Kinase 1 Inhibitor, PF543, Mitigates Pulmonary Fibrosis by Reducing Lung Epithelial Cell mtDNA Damage and Recruitment of Fibrogenic Monocytes. International Journal of Molecular Sciences. 2020; 21(16):5595. https://doi.org/10.3390/ijms21165595
Chicago/Turabian StyleCheresh, Paul, Seok-Jo Kim, Long Shuang Huang, Satoshi Watanabe, Nikita Joshi, Kinola J.N. Williams, Monica Chi, Ziyan Lu, Anantha Harijith, Anjana Yeldandi, and et al. 2020. "The Sphingosine Kinase 1 Inhibitor, PF543, Mitigates Pulmonary Fibrosis by Reducing Lung Epithelial Cell mtDNA Damage and Recruitment of Fibrogenic Monocytes" International Journal of Molecular Sciences 21, no. 16: 5595. https://doi.org/10.3390/ijms21165595