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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

Hepatic injury and CA II expression and activity after cold ischemia and normothermic reperfusion.(A) Hepatic injury measured as AST and ALT levels after 2h-normotermic ex-vivo perfusion. CA II supplementation of IGL-1 attenuated AST/ALT levels, in contrast to the IGL-1 group. (B) Hematoxylin and eosin staining. The IGL-1 group presented increased hepatic damage compared to the control group (Ctr 2), while CA II addition to IGL-1 diminished hepatic damage compared to the IGL-1 group; (C) CA II protein expression by Western blot analyses. CA II expression was increased in the IGL-1+CAII group compared to Ctr 2 and IGL-1 groups; (D) CA II activity levels in fatty livers after 2h-normothermic reperfusion. CA II activity levels decreased in livers preserved in IGL-1 and IGL-1+CAII groups when compared to the control group (Ctr 2); ATP quantitation: ATP levels was partially restored in liver preserved in IGL-1 supplemented with CA II. Ctr 2: Livers flushed and perfused ex-vivo without cold preservation; IGL-1: liver preserved in IGL-1 solution (4°C, 24 h) and then 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.g002: Hepatic injury and CA II expression and activity after cold ischemia and normothermic reperfusion.(A) Hepatic injury measured as AST and ALT levels after 2h-normotermic ex-vivo perfusion. CA II supplementation of IGL-1 attenuated AST/ALT levels, in contrast to the IGL-1 group. (B) Hematoxylin and eosin staining. The IGL-1 group presented increased hepatic damage compared to the control group (Ctr 2), while CA II addition to IGL-1 diminished hepatic damage compared to the IGL-1 group; (C) CA II protein expression by Western blot analyses. CA II expression was increased in the IGL-1+CAII group compared to Ctr 2 and IGL-1 groups; (D) CA II activity levels in fatty livers after 2h-normothermic reperfusion. CA II activity levels decreased in livers preserved in IGL-1 and IGL-1+CAII groups when compared to the control group (Ctr 2); ATP quantitation: ATP levels was partially restored in liver preserved in IGL-1 supplemented with CA II. Ctr 2: Livers flushed and perfused ex-vivo without cold preservation; IGL-1: liver preserved in IGL-1 solution (4°C, 24 h) and then 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: Addition of CA II to IGL-1 solution significantly decreased AST/ALT levels after 2 hours of normothermic perfusion when compared to IGL-1 group (Fig 2A).


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)

Hepatic injury and CA II expression and activity after cold ischemia and normothermic reperfusion.(A) Hepatic injury measured as AST and ALT levels after 2h-normotermic ex-vivo perfusion. CA II supplementation of IGL-1 attenuated AST/ALT levels, in contrast to the IGL-1 group. (B) Hematoxylin and eosin staining. The IGL-1 group presented increased hepatic damage compared to the control group (Ctr 2), while CA II addition to IGL-1 diminished hepatic damage compared to the IGL-1 group; (C) CA II protein expression by Western blot analyses. CA II expression was increased in the IGL-1+CAII group compared to Ctr 2 and IGL-1 groups; (D) CA II activity levels in fatty livers after 2h-normothermic reperfusion. CA II activity levels decreased in livers preserved in IGL-1 and IGL-1+CAII groups when compared to the control group (Ctr 2); ATP quantitation: ATP levels was partially restored in liver preserved in IGL-1 supplemented with CA II. Ctr 2: Livers flushed and perfused ex-vivo without cold preservation; IGL-1: liver preserved in IGL-1 solution (4°C, 24 h) and then 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.g002: Hepatic injury and CA II expression and activity after cold ischemia and normothermic reperfusion.(A) Hepatic injury measured as AST and ALT levels after 2h-normotermic ex-vivo perfusion. CA II supplementation of IGL-1 attenuated AST/ALT levels, in contrast to the IGL-1 group. (B) Hematoxylin and eosin staining. The IGL-1 group presented increased hepatic damage compared to the control group (Ctr 2), while CA II addition to IGL-1 diminished hepatic damage compared to the IGL-1 group; (C) CA II protein expression by Western blot analyses. CA II expression was increased in the IGL-1+CAII group compared to Ctr 2 and IGL-1 groups; (D) CA II activity levels in fatty livers after 2h-normothermic reperfusion. CA II activity levels decreased in livers preserved in IGL-1 and IGL-1+CAII groups when compared to the control group (Ctr 2); ATP quantitation: ATP levels was partially restored in liver preserved in IGL-1 supplemented with CA II. Ctr 2: Livers flushed and perfused ex-vivo without cold preservation; IGL-1: liver preserved in IGL-1 solution (4°C, 24 h) and then 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: Addition of CA II to IGL-1 solution significantly decreased AST/ALT levels after 2 hours of normothermic perfusion when compared to IGL-1 group (Fig 2A).

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