N-acetylglucosamine-1-Phosphate Transferase Suppresses Lysosomal Hydrolases in Dysfunctional Osteoclasts: A Potential Mechanism for Vascular Calcification
Abstract
:1. Introduction
2. Materials and Methods
2.1. Human Carotid Arteries
2.2. Immunohistochemistry and Immunofluorescence for Human Tissues
2.3. CD14-Immunoreactive Monocyte Isolation and Osteoclast Differentiation
2.4. siRNA Transfection Experiments
2.5. Real-Time PCR Analysis
2.6. Immunofluorescence for Human Cells
2.7. TRAP Staining and Quantification
2.8. Resorption Assay for Osteoclast Functional Analysis
2.9. CTSK Activity Assay
2.10. Statistics
3. Results
3.1. Human Calcified Arteries Contain Enriched GNPTAB around Calcified Areas but Low Levels of Osteoclast Lysosomal Hydrolases
Human Arteries ID | GNPTAB | CD68 | TRAP | CTSK | |
---|---|---|---|---|---|
Non-Calcified Human Carotid Arteries (n = 3) | |||||
NC1-CAA5 | - | + | - | + | |
NC2-CAA8 | - | + | - | + | |
NC3-CAA11 | - | + | - | + | |
Calcified Human Carotid Arteries (n = 6) | |||||
C1-CAA2b | +++ | ++++ | - | + | |
C2-LCE1 | ++ | ++++ | + | + | |
C3-LCE4 | ++++ | ++++ | + | - | |
C4-LCE11 | ++++ | ++++ | + | - | |
C5-RCE7 | + | ++ | - | - | |
C6-RCE10 | ++ | ++++ | + | - |
3.2. GNPTAB Silencing Increases Osteoclastogenesis and Resorption Pits Formed by Functional Osteoclast-Like Cells
3.3. GNPTAB Silencing Increased Secretion of Lysosomal Hydrolases
4. Discussion
5. Conclusions
Supplementary Materials
Acknowledgments
Author Contributions
Conflicts of Interest
References
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Lei, Y.; Iwashita, M.; Choi, J.; Aikawa, M.; Aikawa, E. N-acetylglucosamine-1-Phosphate Transferase Suppresses Lysosomal Hydrolases in Dysfunctional Osteoclasts: A Potential Mechanism for Vascular Calcification. J. Cardiovasc. Dev. Dis. 2015, 2, 31-47. https://doi.org/10.3390/jcdd2020031
Lei Y, Iwashita M, Choi J, Aikawa M, Aikawa E. N-acetylglucosamine-1-Phosphate Transferase Suppresses Lysosomal Hydrolases in Dysfunctional Osteoclasts: A Potential Mechanism for Vascular Calcification. Journal of Cardiovascular Development and Disease. 2015; 2(2):31-47. https://doi.org/10.3390/jcdd2020031
Chicago/Turabian StyleLei, Yang, Masaya Iwashita, Jung Choi, Masanori Aikawa, and Elena Aikawa. 2015. "N-acetylglucosamine-1-Phosphate Transferase Suppresses Lysosomal Hydrolases in Dysfunctional Osteoclasts: A Potential Mechanism for Vascular Calcification" Journal of Cardiovascular Development and Disease 2, no. 2: 31-47. https://doi.org/10.3390/jcdd2020031