Vitamin C Protects Chondrocytes against Monosodium Iodoacetate-Induced Osteoarthritis by Multiple Pathways
Abstract
:1. Introduction
2. Results
2.1. Cell Growth Inhibition
2.2. Oxidative Stress
2.3. Apoptosis, Cell Cycle Progress, and Apoptosis-Related Proteins
2.4. Proteoglycan Loss
2.5. Expression of Pro-Inflammation Cytokine and MMP
2.6. Articular Cartilage Loss of the MIA-Induced OA in Rats
3. Discussion
4. Materials and Methods
4.1. Reagents and Antibodies
4.2. Cell Culture
4.3. Cell Viability Assay and Morphology
4.4. Measurement of Reactive Oxygen Species (ROS)
4.5. Detection of Apoptosis and Cell Cycle Progress
4.6. Western Blot Analysis
4.7. Glycosaminoglycans Staining
4.8. Quantitative Real-Time PCR Analysis
4.9. Animals and Treatments
4.10. Histopathology of Joint Tissues: Safranin O and Fast Green Staining
4.11. Serum Biomarker Measurements
4.12. Statistical Analysis
5. Conclusions
Acknowledgments
Author Contributions
Conflicts of Interest
Abbreviations
OA | Osteoarthritis |
MIA | Monosodium Iodoacetate |
MMPs | Matrix Metalloproteinases |
IL | Interleukin |
TNF-α | Tumor Necrosis Factor-alpha |
Vit. C | Vitamin C |
ROS | Reactive Oxygen Species |
ADAMTS | A Disintegrin And Metalloproteinase with Thrombospondin Motifs |
MAPKs | Mitogen-Activated Protein Kinases |
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Primer Name | NCBI Reference Sequence | Primer Sequence (5′→3′) |
---|---|---|
β-actin | NM_001101.3 | F: ATCGGCGGCTCCATCCTG |
R: ACTCGTCATACTCCTGCTTGC | ||
MMP-1 | NM_002421.3 | F: AGATGTGGAGTGCCTGATGTG |
R: CTTGACCCTCAGAGACCTTGG | ||
MMP-3 | NM_002422.3 | F: CCACTCTATCACTCACTCACAG |
R: GACAGCATCAAAGGACAAAGC | ||
MMP-9 | NM_004994.2 | F: CTGGTCCTGGTGCTCCTG |
R: TGCCTGTCGGTGAGATTGG | ||
MMP-13 | NM_002427.3 | F: GACCCTGGAGCACTCATGTTTC |
R: TCCTCGGAGACTGGTAATGGC | ||
IL-1β | NM_000576.2 | F: TGATGGCTTATTACAGTGGCAATG |
R: GTAGTGGTGGTCGGAGATTCG | ||
IL-6 | NM_000600.4 | F: ACCCCCAATAAATATAGGACTGGA |
R: GAGAAGGCAACTGGACCGAA | ||
TNF-α | NM_000594.3 | F: TCAGCAAGGACAGCAGAGGAC |
R: GGAGCCGTGGGTCAGTATGTG | ||
IL-17A | NM_002190.2 | F: GGCTGGAGAAGATACTGGTGTC |
R: AGGCTGTCTTTGAAGGATGAGG |
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Chiu, P.-R.; Hu, Y.-C.; Huang, T.-C.; Hsieh, B.-S.; Yeh, J.-P.; Cheng, H.-L.; Huang, L.-W.; Chang, K.-L. Vitamin C Protects Chondrocytes against Monosodium Iodoacetate-Induced Osteoarthritis by Multiple Pathways. Int. J. Mol. Sci. 2017, 18, 38. https://doi.org/10.3390/ijms18010038
Chiu P-R, Hu Y-C, Huang T-C, Hsieh B-S, Yeh J-P, Cheng H-L, Huang L-W, Chang K-L. Vitamin C Protects Chondrocytes against Monosodium Iodoacetate-Induced Osteoarthritis by Multiple Pathways. International Journal of Molecular Sciences. 2017; 18(1):38. https://doi.org/10.3390/ijms18010038
Chicago/Turabian StyleChiu, Pu-Rong, Yu-Chen Hu, Tzu-Ching Huang, Bau-Shan Hsieh, Jou-Pei Yeh, Hsiao-Ling Cheng, Li-Wen Huang, and Kee-Lung Chang. 2017. "Vitamin C Protects Chondrocytes against Monosodium Iodoacetate-Induced Osteoarthritis by Multiple Pathways" International Journal of Molecular Sciences 18, no. 1: 38. https://doi.org/10.3390/ijms18010038