Homocysteine Disrupts Balance between MMP-9 and Its Tissue Inhibitor in Diabetic Retinopathy: The Role of DNA Methylation
Abstract
:1. Introduction
2. Results
2.1. In Vitro: Retinal Endothelial Cells in Culture
2.2. In Vivo Model: Rat Retinal Microvasculature
3. Discussion
4. Materials and Methods
4.1. Diabetic Rats
4.2. Retinal Microvessels
4.3. Homocysteine Levels
4.4. Gene Transcripts
4.5. Western Blotting
4.6. Activities of Enzymes
4.7. Immunofluorescence
4.8. Co-Immunoprecipitation
4.9. Reactive Oxygen Species
4.10. Apoptosis of Retinal Endothelial Cells
4.11. Methylated/Hydroxymethylated Cytosine
4.12. Chromatin Immunoprecipitation (ChIP)
4.13. Statistical Analysis
Author Contributions
Funding
Acknowledgments
Conflicts of Interest
References
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Gene | Sequence |
---|---|
Human | |
MMP-9 | Fwd- CACTGTCCACCCCTCAGAGC |
Rev- GCCACTTGTCGGCGATAAGG | |
Timp1 | Fwd- GAAGAGCCTGAACCACAGGT |
Rev- CGGGGAGGAGATGTAGCAC | |
MMP-9 promoter | Fwd- GAGTCAGCACTTGCCTGTCA |
Rev- CTGCTGTTGTGGGGGCTTTA | |
Timp1 promoter | Fwd- CTGATGGTGGGTGGATGAGTAATG |
Rev- GCCTGAGCGCTAGAGGATAAATG | |
β-Actin | Fwd- AGCCTCGCCTTTGCCGATCCG |
Rev- TCTCTTGCTCTGGGCCTCGTCG | |
Rat | |
MMP-9 | Fwd- GAAGCAGCTGTCCCTGCCCC |
Rev- GATGGTGCCACTTGAGGTCG | |
Timp1 | Fwd- GTTTCCGGTTCGCCTACACC |
Rev- GGGCTCAGATTATGCCAGGG | |
MMP-9 promoter | Fwd- GACTGTGGGCAGGGCATAAA |
Rev- GGGTGAAGCAGAATTTGCGG | |
Timp1 promoter | Fwd- GCAGTGGGTGGATGAGTAAT |
Rev- GACTGGCGGTGGAGAATA AA | |
β-Actin | Fwd- TCTGTGTGGATTGGTGGCTCTA |
Rev- AAC AGTCCGCCTAGAAGCATTTG |
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Mohammad, G.; Kowluru, R.A. Homocysteine Disrupts Balance between MMP-9 and Its Tissue Inhibitor in Diabetic Retinopathy: The Role of DNA Methylation. Int. J. Mol. Sci. 2020, 21, 1771. https://doi.org/10.3390/ijms21051771
Mohammad G, Kowluru RA. Homocysteine Disrupts Balance between MMP-9 and Its Tissue Inhibitor in Diabetic Retinopathy: The Role of DNA Methylation. International Journal of Molecular Sciences. 2020; 21(5):1771. https://doi.org/10.3390/ijms21051771
Chicago/Turabian StyleMohammad, Ghulam, and Renu A. Kowluru. 2020. "Homocysteine Disrupts Balance between MMP-9 and Its Tissue Inhibitor in Diabetic Retinopathy: The Role of DNA Methylation" International Journal of Molecular Sciences 21, no. 5: 1771. https://doi.org/10.3390/ijms21051771