VEGF-Independent Activation of Müller Cells by the Vitreous from Proliferative Diabetic Retinopathy Patients
Abstract
:1. Introduction
2. Results
2.1. MIO-M1 Müller Cells Are Activated by PDR Vitreous
2.2. PDR Vitreous-Induced Activation of Müller Cells Is Independent from VEGF
3. Discussion
4. Materials and Methods
4.1. Reagents
4.2. Human Vitreous Fluid Samples
4.3. Cell Cultures
4.4. MIO-M1 Proliferation Assay
4.5. MIO-M1 Wound Healing Assay
4.6. Western Blot Analysis
4.7. RT-PCR Analyses
4.8. MIO-M1 Immunofluorescence Analysis
4.9. Statistical Analysis
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
Abbreviations
ACTA2 | actin α 2 |
ERM | epiretinal membrane |
FGF2 | basic fibroblast growth factor |
FGFR | fibroblast growth factor receptor |
GAPDH | glyceraldehyde-3-phosphate dehydrogenase |
HUVECs | human umbilical vein endothelial cells |
IL | interleukin |
INFγ | interferon γ |
MCP1 | monocyte chemoattractant protein 1 |
NLRP3 | nucleotide-binding oligomerization domain (NOD), leucine-rich repeat (LRR)-containing proteins 3 |
PDGF | platelet derived growth factor |
PDR | proliferative diabetic retinopathy |
RLBP1 | retinaldehyde binding protein 1 |
S100A4 | S100 calcium-binding protein A4 |
TNFα | tumor necrosis factor α |
VEGF-A | vascular endothelial growth factor-A |
VEGFR | vascular endothelial growth factor receptor |
VIM | vimentin |
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Patients/Eyes | 39/42 |
---|---|
Clinical features | |
Gender (male/female) | 28/11 |
Age (years) | 65 ± 10 |
Type 1/type 2 diabetes | 4/35 |
Duration of diabetes (years) | 21 ± 6 |
Oral hypoglycemic drug treatment | 10/39 |
Insulin treatment | 10/39 |
Oral hypoglycemic drug + insulin treatment | 19/39 |
Glycaemia (mg/dL) | 161 ± 56 |
HbA1c (%) | 7.9 ± 1.1 |
Neuropathy | 6/39 |
Nephropathy | 13/39 |
Cardiopathy | 15/39 |
Hypertension | 37/39 |
Dyslipidemia | 23/39 |
Triglycerides (mg/dL) | 120 ± 54 |
Cholesterol (mg/dL) | 153 ± 44 |
Creatinine (mg/dL) | 1.4 ± 0.7 |
Hemoglobin (g/dL) | 13.1 ± 1.6 |
Ophthalmic features | |
PDR | 42/42 |
PDR with vitreous hemorrhage | 19/42 |
PDR with macular edema | 19/42 |
PDR with ERM | 31/38 |
Ocular therapies | |
Intravitreal injection of anti-VEGF blocker | 29/42 |
Panretinal laser photocoagulation | 32/42 |
Gene | Forward | Reverse |
---|---|---|
ACTA2 | 5′-AATGGCTCTGGGCTCTGTAA-3′ | 5′-TTTTGCTCTGTGCTTCGTCA-3′ |
FGFR1 | 5′-GGGCTGGAATACTGCTACAA-3′ | 5′-GCCAAAGTCTGCTATCTTCATC-3′ |
FGFR2 | 5′-GGATAACAACACGCCTCTCTT-3′ | 5′-GCCCAAAGCAACCTTCTC-3′ |
FGFR3 | 5′-TGGTGTCCTGTGCCTACC-3′ | 5′-CCGTTGGTCGTCTTCTTGT-3′ |
FGFR4 | 5′-AACCGCATTGGAGGCATT-3′ | 5′-TCTACCAGGCAGGTGTATGT-3′ |
GAPDH | 5′-GAAGGTCGGAGTCAACGGATT-3′ | 5′-TGACGGTGCCATGGAATTTG-3′ |
IL1β | 5′-GTGGCAATGAGGATGACTTG-3′ | 5′-GTGGTGGTCGGAGATTCGTA-3′ |
IL6 | 5′-TGTGTGGGTCTGTTGTAGGG-3′ | 5′-CCCGTGCAATATCTAGGAAAA-3′ |
IL8 | 5′-TGTGTGGGTCTGTTGTAGGG-3′ | 5′-CCCGTGCAATATCTAGGAAAA-3′ |
INFγ | 5′-GCAGGTCATTCAGATGTAGCGG-3′ | 5′-CCACACTCTTTTGGATGCTCTGG-3′ |
MCP1 | 5′-CTCAGCCAGATGCAATCAA-3′ | 5′-CACTTCTGCTTGGGGTCA-3′ |
NLRP3 | 5′-GGACTGAAGCACCTGTTGTGCA-3′ | 5′-TCCTGAGTCTCCCAAGGCATTC-3′ |
RLBP1 | 5′-GCTGCTGGAGAATGAGGAAA-3′ | 5′-TGGTGGATGAAGTGGATGG-3′ |
S100A4 | 5′-CCTGGATGTGATGGTGTCC-3′ | 5′-TCGTTGTCCCTGTTGCTGT-3′ |
TNFα | 5′-TGCTTGTTCCTCAGCCTCTT-3′ | 5′-GCTTGTCACTCGGGGTTC-3′ |
VEGF-A | 5′-AATCGAGACCCTGGTGGAC-3′ | 5′-GGTGAGGTTTGATCCGCATA-3′ |
VEGFR1 | 5′-AGCAGTTCCACCACTTTAGA-3′ | 5′-GAACTTTCCACAGAGCCCTT-3′ |
VEGFR2 | 5′-GGAAATGACACTGGAGCCTA-3′ | 5′-TTTGAAATGGACCCGAGACA-3′ |
VEGFR3 | 5′-CAACGACCTACAAAGGCTCT-3′ | 5′-GTAAAACACCTGGCCTCCTC-3′ |
VIM | 5′-CGCCAGATGCGTGAAATG-3′ | 5′-ACCAGAGGGAGTGAATCCAGA-3′ |
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Rezzola, S.; Guerra, J.; Krishna Chandran, A.M.; Loda, A.; Cancarini, A.; Sacristani, P.; Semeraro, F.; Presta, M. VEGF-Independent Activation of Müller Cells by the Vitreous from Proliferative Diabetic Retinopathy Patients. Int. J. Mol. Sci. 2021, 22, 2179. https://doi.org/10.3390/ijms22042179
Rezzola S, Guerra J, Krishna Chandran AM, Loda A, Cancarini A, Sacristani P, Semeraro F, Presta M. VEGF-Independent Activation of Müller Cells by the Vitreous from Proliferative Diabetic Retinopathy Patients. International Journal of Molecular Sciences. 2021; 22(4):2179. https://doi.org/10.3390/ijms22042179
Chicago/Turabian StyleRezzola, Sara, Jessica Guerra, Adwaid Manu Krishna Chandran, Alessandra Loda, Anna Cancarini, Piergiuseppe Sacristani, Francesco Semeraro, and Marco Presta. 2021. "VEGF-Independent Activation of Müller Cells by the Vitreous from Proliferative Diabetic Retinopathy Patients" International Journal of Molecular Sciences 22, no. 4: 2179. https://doi.org/10.3390/ijms22042179