Activated Protein C Ameliorates Tubular Mitochondrial Reactive Oxygen Species and Inflammation in Diabetic Kidney Disease
Abstract
:1. Introduction
2. Materials and Methods
2.1. Reagents
2.2. Mice
2.3. Determination of Albuminuria
2.4. Cell Culture
2.5. RNA Expression Profiling
2.6. Flow Cytometry
2.7. Immunoblotting
2.8. Quantitative Polymerase Chain Reaction (qPCR)
2.9. Total DNA Isolation
2.10. ATP Level Quantification
2.11. Histochemistry
2.12. Immunofluorescence
2.13. Transmission Electron Microscopy
3. Results
3.1. aPC Regulates Pathways Related to Metabolism and Oxidoreductase Activity in DKD
3.2. aPC Protects against Mitochondrial Dysfunction in DKD
3.3. aPC Reduces Tubular ROS in Diabetic Nephropathy
3.4. aPC Reduces Inflammasome Activation in Diabetic Nephropathy
4. Discussion
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
Abbreviations
8-oxo-dG | 8-Oxo-2’-deoxyguanosine |
aPC | Activated protein C |
ANOVA | Analysis of variance |
ASC | Apoptosis-associated speck-like protein containing a CARD |
BUMPT | Boston university mouse proximal tubular cells |
DAPI | 4′,6-diamidino-2-phenylindole |
DAVID | Database for Annotation, Visualization, and Integrated Discovery |
DKD | Diabetic kidney disease |
DM | Diabetes mellitus |
EPCR | Endothelial protein C receptor |
FACS | Fluorescence-activated cell sorting |
GBM | Glomerular basement membrane |
APChigh | Hyperactivatable protein C |
IL-1β | Interleukin 1 beta |
KEGG | Kyoto Encyclopedia of Genes and Genomes |
MFI | Mean Fluorescence Intensity |
MTS | Masson’s trichrome stain |
NF-κB | Nuclear factor kappa-light-chain-enhancer of activated B cells |
NRLP3 | NOD-, LRR- and pyrin domain-containing protein 3 |
NO | Nitric oxide |
PAS | Periodic acid–Schiff staining |
PGC-1α | Peroxisome proliferator-activated receptor gamma coactivator 1-alpha |
qRT-PCR | Quantitative real time polymerase chain reaction |
ROS | Reactive oxygen species |
SEM | Standard error mean |
STZ | Streptozotocin |
UCAR | Urine albumin-creatinine ratio |
VDAC | Voltage-dependent anion-selective channel |
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Rana, R.; Manoharan, J.; Gupta, A.; Gupta, D.; Elwakiel, A.; Khawaja, H.; Fatima, S.; Zimmermann, S.; Singh, K.; Ambreen, S.; et al. Activated Protein C Ameliorates Tubular Mitochondrial Reactive Oxygen Species and Inflammation in Diabetic Kidney Disease. Nutrients 2022, 14, 3138. https://doi.org/10.3390/nu14153138
Rana R, Manoharan J, Gupta A, Gupta D, Elwakiel A, Khawaja H, Fatima S, Zimmermann S, Singh K, Ambreen S, et al. Activated Protein C Ameliorates Tubular Mitochondrial Reactive Oxygen Species and Inflammation in Diabetic Kidney Disease. Nutrients. 2022; 14(15):3138. https://doi.org/10.3390/nu14153138
Chicago/Turabian StyleRana, Rajiv, Jayakumar Manoharan, Anubhuti Gupta, Dheerendra Gupta, Ahmed Elwakiel, Hamzah Khawaja, Sameen Fatima, Silke Zimmermann, Kunal Singh, Saira Ambreen, and et al. 2022. "Activated Protein C Ameliorates Tubular Mitochondrial Reactive Oxygen Species and Inflammation in Diabetic Kidney Disease" Nutrients 14, no. 15: 3138. https://doi.org/10.3390/nu14153138