Activated Alpha 2-Macroglobulin Is a Novel Mediator of Mesangial Cell Profibrotic Signaling in Diabetic Kidney Disease
Abstract
:1. Introduction
2. Materials and Methods
2.1. Cell Culture
2.2. Protein Extraction and Immunoblotting
2.3. qPCR
2.4. Experimental Animals and Tissue Processing
2.5. Transfection
2.6. Intracellular Calcium Assay
2.7. Cell Surface Protein Isolation
2.8. TGFβ1 ELISA
2.9. Recovery of α2M* from Urine Samples
2.10. Patient Cohort
2.11. Statistical Analysis
3. Results
3.1. α2M Is Increased and Activated by HG in MCs and in Diabetic Kidneys
3.2. Inhibition of α2M/α2M* Inhibits HG-Induced Profibrotic Responses by MCs
3.3. LRP1 Is Not Involved in HG-Induced Profibrotic Responses in MCs
3.4. Increased Matrix Synthesis with csGRP78 Overexpression Requires α2M*
3.5. α2M* Regulates TGFβ1 Production by HG in MCs
3.6. Urinary α2M* Is Associated with Urinary TGFβ1 in Individuals with Overt DKD
4. Discussion
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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Number | 18 |
Male, n (%) | 16 (89) |
Age (years) | 70 ± 8 |
Baseline eGFR (mL/min/1.73 m²) | 25 ± 10 |
eGFR decline rate (mL/min/1.73 m²/year) | −2 ± 2 |
Follow-up period (year) | 2.3 ± 0.6 |
Protein/creatinine ratio (g/g) | 1.3 ± 0.5 |
SBP (mmHg) | 142 ± 14 |
DBP (mmHg) | 66 ± 8 |
RASB use, n (%) | 17 (94) |
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Trink, J.; Li, R.; Palarasah, Y.; Troyanov, S.; Andersen, T.E.; Sidelmann, J.J.; Inman, M.D.; Pizzo, S.V.; Gao, B.; Krepinsky, J.C. Activated Alpha 2-Macroglobulin Is a Novel Mediator of Mesangial Cell Profibrotic Signaling in Diabetic Kidney Disease. Biomedicines 2021, 9, 1112. https://doi.org/10.3390/biomedicines9091112
Trink J, Li R, Palarasah Y, Troyanov S, Andersen TE, Sidelmann JJ, Inman MD, Pizzo SV, Gao B, Krepinsky JC. Activated Alpha 2-Macroglobulin Is a Novel Mediator of Mesangial Cell Profibrotic Signaling in Diabetic Kidney Disease. Biomedicines. 2021; 9(9):1112. https://doi.org/10.3390/biomedicines9091112
Chicago/Turabian StyleTrink, Jackie, Renzhong Li, Yaseelan Palarasah, Stéphan Troyanov, Thomas E. Andersen, Johannes J. Sidelmann, Mark D. Inman, Salvatore V. Pizzo, Bo Gao, and Joan C. Krepinsky. 2021. "Activated Alpha 2-Macroglobulin Is a Novel Mediator of Mesangial Cell Profibrotic Signaling in Diabetic Kidney Disease" Biomedicines 9, no. 9: 1112. https://doi.org/10.3390/biomedicines9091112