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Carbonic Anhydrase Protects Fatty Liver Grafts against Ischemic Reperfusion Damage.

Bejaoui M, Pantazi E, De Luca V, Panisello A, Folch-Puy E, Hotter G, Capasso C, T Supuran C, Roselló-Catafau J, Rosselló-Catafau J - PLoS ONE (2015)

Bottom Line: In all cases, liver injury, CA II protein concentration, CA II mRNA levels and CA II activity were determined.In the case of reperfusion, CA II protection was associated with better function, AMPK activation and the prevention of ERS and MAPKs activation.Interestingly, CA II supplementation was not associated with enhanced CO2 hydration.

View Article: PubMed Central - PubMed

Affiliation: Department of Experimental Pathology, Institute of Biomedical Research of Barcelona-Spanish National Research Council (IIBB-CSIC), IDIBAPS, Barcelona, Spain.

ABSTRACT
Carbonic anhydrases (CAs) are ubiquitous metalloenzymes that catalyze the reversible hydration of carbon dioxide to bicarbonate and a proton. CAs are involved in numerous physiological and pathological processes, including acid-base homeostasis, electrolyte balance, oxygen delivery to tissues and nitric oxide generation. Given that these processes are found to be dysregulated during ischemia reperfusion injury (IRI), and taking into account the high vulnerability of steatotic livers to preservation injury, we hypothesized a new role for CA as a pharmacological agent able to protect against ischemic damage. Two different aspects of the role of CA II in fatty liver grafts preservation were evaluated: 1) the effect of its addition to Institut Georges Lopez (IGL-1) storage solution after cold ischemia; 2) and after 24h of cold storage followed by two hours of normothermic ex-vivo perfusion. In all cases, liver injury, CA II protein concentration, CA II mRNA levels and CA II activity were determined. In case of the ex-vivo perfusion, we further assessed liver function (bile production, bromosulfophthalein clearance) and Western blot analysis of phosphorylated adenosine monophosphate activated protein kinase (AMPK), mitogen activated protein kinases family (MAPKs) and endoplasmic reticulum stress (ERS) parameters (GRP78, PERK, IRE, eIF2α and ATF6). We found that CA II was downregulated after cold ischemia. The addition of bovine CA II to IGL-1 preservation solution efficiently protected steatotic liver against cold IRI. In the case of reperfusion, CA II protection was associated with better function, AMPK activation and the prevention of ERS and MAPKs activation. Interestingly, CA II supplementation was not associated with enhanced CO2 hydration. The results suggest that CA II modulation may be a promising target for fatty liver graft preservation.

No MeSH data available.


Related in: MedlinePlus

Effects of CA II addition to IGL-1 solution in liver apoptosis.Liver apoptosis, measured as cleaved caspase 9/ total caspases 9 (A), cleaved caspase 3/ total caspase 3 (B) and TUNEL assay (C) in steatotic livers after 120 min of normothermic “ex vivo” perfusion. CA II addition to IGL-1 solution reduced activation of liver caspases 3 and 9 and diminished TUNEL-positive cells. Ctr 2: Liver flushed and perfused “ex-vivo” without cold preservation; IGL-1: liver preserved in IGL-1 solution (4°C, 24 h) and subjected to 2h- normothermic “ex vivo” perfusion. IGL-1+CAII: liver preserved in IGL-1 solution (4°C, 24 h) enriched with CA II and subjected to 2h-normothermic “ex vivo” perfusion. * p < 0.05 vs Ctr 2; # p < 0.05 vs IGL-1.
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pone.0134499.g006: Effects of CA II addition to IGL-1 solution in liver apoptosis.Liver apoptosis, measured as cleaved caspase 9/ total caspases 9 (A), cleaved caspase 3/ total caspase 3 (B) and TUNEL assay (C) in steatotic livers after 120 min of normothermic “ex vivo” perfusion. CA II addition to IGL-1 solution reduced activation of liver caspases 3 and 9 and diminished TUNEL-positive cells. Ctr 2: Liver flushed and perfused “ex-vivo” without cold preservation; IGL-1: liver preserved in IGL-1 solution (4°C, 24 h) and subjected to 2h- normothermic “ex vivo” perfusion. IGL-1+CAII: liver preserved in IGL-1 solution (4°C, 24 h) enriched with CA II and subjected to 2h-normothermic “ex vivo” perfusion. * p < 0.05 vs Ctr 2; # p < 0.05 vs IGL-1.

Mentions: IRI is also associated with the initiation of apoptotic pathways [25]. In our conditions, we observed that preservation of fatty livers in IGL-1 solution supplemented by CA II caused a substantial decrease in liver apoptosis, as reflected by reductions in cleaved caspases 9 and 3 protein expression and TUNEL staining (Fig 6A, 6B and 6D respectively).


Carbonic Anhydrase Protects Fatty Liver Grafts against Ischemic Reperfusion Damage.

Bejaoui M, Pantazi E, De Luca V, Panisello A, Folch-Puy E, Hotter G, Capasso C, T Supuran C, Roselló-Catafau J, Rosselló-Catafau J - PLoS ONE (2015)

Effects of CA II addition to IGL-1 solution in liver apoptosis.Liver apoptosis, measured as cleaved caspase 9/ total caspases 9 (A), cleaved caspase 3/ total caspase 3 (B) and TUNEL assay (C) in steatotic livers after 120 min of normothermic “ex vivo” perfusion. CA II addition to IGL-1 solution reduced activation of liver caspases 3 and 9 and diminished TUNEL-positive cells. Ctr 2: Liver flushed and perfused “ex-vivo” without cold preservation; IGL-1: liver preserved in IGL-1 solution (4°C, 24 h) and subjected to 2h- normothermic “ex vivo” perfusion. IGL-1+CAII: liver preserved in IGL-1 solution (4°C, 24 h) enriched with CA II and subjected to 2h-normothermic “ex vivo” perfusion. * p < 0.05 vs Ctr 2; # p < 0.05 vs IGL-1.
© Copyright Policy
Related In: Results  -  Collection

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Show All Figures
getmorefigures.php?uid=PMC4520486&req=5

pone.0134499.g006: Effects of CA II addition to IGL-1 solution in liver apoptosis.Liver apoptosis, measured as cleaved caspase 9/ total caspases 9 (A), cleaved caspase 3/ total caspase 3 (B) and TUNEL assay (C) in steatotic livers after 120 min of normothermic “ex vivo” perfusion. CA II addition to IGL-1 solution reduced activation of liver caspases 3 and 9 and diminished TUNEL-positive cells. Ctr 2: Liver flushed and perfused “ex-vivo” without cold preservation; IGL-1: liver preserved in IGL-1 solution (4°C, 24 h) and subjected to 2h- normothermic “ex vivo” perfusion. IGL-1+CAII: liver preserved in IGL-1 solution (4°C, 24 h) enriched with CA II and subjected to 2h-normothermic “ex vivo” perfusion. * p < 0.05 vs Ctr 2; # p < 0.05 vs IGL-1.
Mentions: IRI is also associated with the initiation of apoptotic pathways [25]. In our conditions, we observed that preservation of fatty livers in IGL-1 solution supplemented by CA II caused a substantial decrease in liver apoptosis, as reflected by reductions in cleaved caspases 9 and 3 protein expression and TUNEL staining (Fig 6A, 6B and 6D respectively).

Bottom Line: In all cases, liver injury, CA II protein concentration, CA II mRNA levels and CA II activity were determined.In the case of reperfusion, CA II protection was associated with better function, AMPK activation and the prevention of ERS and MAPKs activation.Interestingly, CA II supplementation was not associated with enhanced CO2 hydration.

View Article: PubMed Central - PubMed

Affiliation: Department of Experimental Pathology, Institute of Biomedical Research of Barcelona-Spanish National Research Council (IIBB-CSIC), IDIBAPS, Barcelona, Spain.

ABSTRACT
Carbonic anhydrases (CAs) are ubiquitous metalloenzymes that catalyze the reversible hydration of carbon dioxide to bicarbonate and a proton. CAs are involved in numerous physiological and pathological processes, including acid-base homeostasis, electrolyte balance, oxygen delivery to tissues and nitric oxide generation. Given that these processes are found to be dysregulated during ischemia reperfusion injury (IRI), and taking into account the high vulnerability of steatotic livers to preservation injury, we hypothesized a new role for CA as a pharmacological agent able to protect against ischemic damage. Two different aspects of the role of CA II in fatty liver grafts preservation were evaluated: 1) the effect of its addition to Institut Georges Lopez (IGL-1) storage solution after cold ischemia; 2) and after 24h of cold storage followed by two hours of normothermic ex-vivo perfusion. In all cases, liver injury, CA II protein concentration, CA II mRNA levels and CA II activity were determined. In case of the ex-vivo perfusion, we further assessed liver function (bile production, bromosulfophthalein clearance) and Western blot analysis of phosphorylated adenosine monophosphate activated protein kinase (AMPK), mitogen activated protein kinases family (MAPKs) and endoplasmic reticulum stress (ERS) parameters (GRP78, PERK, IRE, eIF2α and ATF6). We found that CA II was downregulated after cold ischemia. The addition of bovine CA II to IGL-1 preservation solution efficiently protected steatotic liver against cold IRI. In the case of reperfusion, CA II protection was associated with better function, AMPK activation and the prevention of ERS and MAPKs activation. Interestingly, CA II supplementation was not associated with enhanced CO2 hydration. The results suggest that CA II modulation may be a promising target for fatty liver graft preservation.

No MeSH data available.


Related in: MedlinePlus