Selective Blockade of the Metabotropic Glutamate Receptor mGluR5 Protects Mouse Livers in In Vitro and Ex Vivo Models of Ischemia Reperfusion Injury
Abstract
:1. Introduction
2. Results
2.1. mGluR5 NAMs Protect Rat Hepatocytes from Ischemic Injury Regardless of Their Ability to Deplete ATP
2.1.1. Effects of MPEP and MTEP on Cellular and Mitochondrial ATP Content
2.1.2. MPEP (30 µM) Does Not Alter Mitochondrial Functionality
2.1.3. MPEP, MTEP and Fenobam Protect Rat Hepatocytes from Warm Ischemic Injury
2.1.4. MPEP Prevents ATP Depletion in Hypoxic Hepatocytes
2.2. MPEP Protects Mouse Livers in an Ex Vivo Model of Cold Storage/Reperfusion Injury
2.3. MPEP Protects Mouse Livers in an Ex Vivo Model of Warm Ischemia Reperfusion Injury
3. Discussion
3.1. MPEP Sequesters ATP from Acellular Solution and Cellular Suspensions
3.2. NAMs Protect Hepatocytes from Ischemic Injury, Regardless of Their Ability to Induce ATP Depletion
3.3. The Blockade of mGluR5 Protects Mouse Livers from Cold Ischemia and Warm Ischemia Reperfusion Injury
4. Materials and Methods
4.1. In Vitro Studies
4.1.1. Mitochondrial Isolation and Assays
4.1.2. Hepatocyte Isolation
4.1.3. Warm Ischemia Experiments on Rat Hepatocytes
4.1.4. Measurement of Cell Viability
4.1.5. Measurement of ATP
4.2. Ex Vivo Studies
4.2.1. Cold Ischemia Reperfusion Model
4.2.2. Warm Ischemia Model
4.2.3. Perfusate Analysis
4.2.4. Western Blot Analysis
4.2.5. Statistical Analysis
Acknowledgments
Author Contributions
Conflicts of Interest
Abbreviations
[3H]InsP | [3H]inositolmonophosphate |
ACPD | (1S,3R)-1-aminocyclopentane-1,3-dicarboxylic acid |
ADP | adenosine diphosphate |
DCFH-DA | dichlorodilhydrofluorescein diacetate |
DFB | 3,3′-difluorobenzaldazine |
DMSO | dimethyl sulfoxide |
DTT | 1,4-dithiothreitol |
eNOS | endothelial nitric oxide synthase |
ER | endoplasmic reticulum |
Gluk2 | glutamate receptor, ionotropic kainate 2 |
iNOS | inducible nitric oxide synthase |
KO | knock-out |
PCR | polymerase chain reaction |
PVDF | polyvinylidene difluoride |
SDS | sodium dodecyl sulfate |
TBS | tris-buffered saline |
TNF-α | tumor necrosis factor-α |
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CTRL | MPEP 300 nM | KO | DHPG 100 µM |
---|---|---|---|
2.39 ± 0.34 | 3.52 ± 1.56 | 2.81 ± 0.92 | 1.82 ± 0.87 |
CTRL | MPEP 300 nM | KO | DHPG 100 µM |
---|---|---|---|
2.48 ± 0.55 | 2.90 ± 0.70 | 2.44 ± 0.82 | 2.13 ± 0.77 |
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Ferrigno, A.; Berardo, C.; Di Pasqua, L.G.; Siciliano, V.; Richelmi, P.; Nicoletti, F.; Vairetti, M. Selective Blockade of the Metabotropic Glutamate Receptor mGluR5 Protects Mouse Livers in In Vitro and Ex Vivo Models of Ischemia Reperfusion Injury. Int. J. Mol. Sci. 2018, 19, 314. https://doi.org/10.3390/ijms19020314
Ferrigno A, Berardo C, Di Pasqua LG, Siciliano V, Richelmi P, Nicoletti F, Vairetti M. Selective Blockade of the Metabotropic Glutamate Receptor mGluR5 Protects Mouse Livers in In Vitro and Ex Vivo Models of Ischemia Reperfusion Injury. International Journal of Molecular Sciences. 2018; 19(2):314. https://doi.org/10.3390/ijms19020314
Chicago/Turabian StyleFerrigno, Andrea, Clarissa Berardo, Laura Giuseppina Di Pasqua, Veronica Siciliano, Plinio Richelmi, Ferdinando Nicoletti, and Mariapia Vairetti. 2018. "Selective Blockade of the Metabotropic Glutamate Receptor mGluR5 Protects Mouse Livers in In Vitro and Ex Vivo Models of Ischemia Reperfusion Injury" International Journal of Molecular Sciences 19, no. 2: 314. https://doi.org/10.3390/ijms19020314