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Endothelial Dysfunction in Steatotic Human Donor Livers: A Pilot Study of the Underlying Mechanism During Subnormothermic Machine Perfusion

BACKGROUND: Steatosis is a major risk factor for primary nonfunction in liver transplantations. Steatotic livers recover poorly from ischemia reperfusion injury, in part due to alterations in the microcirculation, although the exact mechanism is unclear. In this study, we tested if there were any al...

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Autores principales: Beijert, Irene, Mert, Safak, Huang, Viola, Karimian, Negin, Geerts, Sharon, Hafiz, Ehab O.A., Markmann, James F., Yeh, Heidi, Porte, Robert J., Uygun, Korkut
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Lippincott Williams & Wilkins 2018
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5959347/
https://www.ncbi.nlm.nih.gov/pubmed/29796416
http://dx.doi.org/10.1097/TXD.0000000000000779
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author Beijert, Irene
Mert, Safak
Huang, Viola
Karimian, Negin
Geerts, Sharon
Hafiz, Ehab O.A.
Markmann, James F.
Yeh, Heidi
Porte, Robert J.
Uygun, Korkut
author_facet Beijert, Irene
Mert, Safak
Huang, Viola
Karimian, Negin
Geerts, Sharon
Hafiz, Ehab O.A.
Markmann, James F.
Yeh, Heidi
Porte, Robert J.
Uygun, Korkut
author_sort Beijert, Irene
collection PubMed
description BACKGROUND: Steatosis is a major risk factor for primary nonfunction in liver transplantations. Steatotic livers recover poorly from ischemia reperfusion injury, in part due to alterations in the microcirculation, although the exact mechanism is unclear. In this study, we tested if there were any alterations in the shear stress sensing Kruppel-like factor 2 (KLF2) and its likely downstream consequences in the ex vivo perfused human liver endothelium, which would imply perturbations in microcirculatory flow in macrosteatotic livers disrupts laminar flow to evaluate if this is a potential therapeutic target for steatotic livers. METHODS: Using a subnormothermic machine perfusion system, 5 macrosteatotic and 4 nonsteatotic human livers were perfused for 3 hours. Flow, resistance, and biochemical profile were monitored. Gene expression levels of nitric oxide synthase 3 (eNOS), KLF2, and thrombomodulin were determined. Nitric oxide (NO) was measured in the perfusion fluid and activation of eNOS was measured with Western blotting. RESULTS: Flow dynamics, injury markers, and bile production were similar in both groups. Kruppel-like factor 2 expression was significantly higher in nonsteatotic livers. Western blotting analyses showed significantly higher levels of activated eNOS in nonsteatotic livers, consistent with an increase in NO production over time. Macrosteatotic livers showed decreased KLF2 upregulation, eNOS activity, and NO production during machine perfusion. CONCLUSIONS: These results indicate a perturbed KLF2 sensing in steatotic livers, which aligns with perturbed microcirculatory state. This may indicate endothelial dysfunction and contribute to poor posttransplantation outcomes in fatty livers, and further studies to confirm by evaluation of flow and testing treatments are warranted.
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spelling pubmed-59593472018-05-24 Endothelial Dysfunction in Steatotic Human Donor Livers: A Pilot Study of the Underlying Mechanism During Subnormothermic Machine Perfusion Beijert, Irene Mert, Safak Huang, Viola Karimian, Negin Geerts, Sharon Hafiz, Ehab O.A. Markmann, James F. Yeh, Heidi Porte, Robert J. Uygun, Korkut Transplant Direct Liver Transplantation BACKGROUND: Steatosis is a major risk factor for primary nonfunction in liver transplantations. Steatotic livers recover poorly from ischemia reperfusion injury, in part due to alterations in the microcirculation, although the exact mechanism is unclear. In this study, we tested if there were any alterations in the shear stress sensing Kruppel-like factor 2 (KLF2) and its likely downstream consequences in the ex vivo perfused human liver endothelium, which would imply perturbations in microcirculatory flow in macrosteatotic livers disrupts laminar flow to evaluate if this is a potential therapeutic target for steatotic livers. METHODS: Using a subnormothermic machine perfusion system, 5 macrosteatotic and 4 nonsteatotic human livers were perfused for 3 hours. Flow, resistance, and biochemical profile were monitored. Gene expression levels of nitric oxide synthase 3 (eNOS), KLF2, and thrombomodulin were determined. Nitric oxide (NO) was measured in the perfusion fluid and activation of eNOS was measured with Western blotting. RESULTS: Flow dynamics, injury markers, and bile production were similar in both groups. Kruppel-like factor 2 expression was significantly higher in nonsteatotic livers. Western blotting analyses showed significantly higher levels of activated eNOS in nonsteatotic livers, consistent with an increase in NO production over time. Macrosteatotic livers showed decreased KLF2 upregulation, eNOS activity, and NO production during machine perfusion. CONCLUSIONS: These results indicate a perturbed KLF2 sensing in steatotic livers, which aligns with perturbed microcirculatory state. This may indicate endothelial dysfunction and contribute to poor posttransplantation outcomes in fatty livers, and further studies to confirm by evaluation of flow and testing treatments are warranted. Lippincott Williams & Wilkins 2018-04-23 /pmc/articles/PMC5959347/ /pubmed/29796416 http://dx.doi.org/10.1097/TXD.0000000000000779 Text en Copyright © 2018 The Author(s). Transplantation Direct. Published by Wolters Kluwer Health, Inc. This is an open-access article distributed under the terms of the Creative Commons Attribution-Non Commercial-No Derivatives License 4.0 (CCBY-NC-ND) (http://creativecommons.org/licenses/by-nc-nd/4.0/) , where it is permissible to download and share the work provided it is properly cited. The work cannot be changed in any way or used commercially without permission from the journal.
spellingShingle Liver Transplantation
Beijert, Irene
Mert, Safak
Huang, Viola
Karimian, Negin
Geerts, Sharon
Hafiz, Ehab O.A.
Markmann, James F.
Yeh, Heidi
Porte, Robert J.
Uygun, Korkut
Endothelial Dysfunction in Steatotic Human Donor Livers: A Pilot Study of the Underlying Mechanism During Subnormothermic Machine Perfusion
title Endothelial Dysfunction in Steatotic Human Donor Livers: A Pilot Study of the Underlying Mechanism During Subnormothermic Machine Perfusion
title_full Endothelial Dysfunction in Steatotic Human Donor Livers: A Pilot Study of the Underlying Mechanism During Subnormothermic Machine Perfusion
title_fullStr Endothelial Dysfunction in Steatotic Human Donor Livers: A Pilot Study of the Underlying Mechanism During Subnormothermic Machine Perfusion
title_full_unstemmed Endothelial Dysfunction in Steatotic Human Donor Livers: A Pilot Study of the Underlying Mechanism During Subnormothermic Machine Perfusion
title_short Endothelial Dysfunction in Steatotic Human Donor Livers: A Pilot Study of the Underlying Mechanism During Subnormothermic Machine Perfusion
title_sort endothelial dysfunction in steatotic human donor livers: a pilot study of the underlying mechanism during subnormothermic machine perfusion
topic Liver Transplantation
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5959347/
https://www.ncbi.nlm.nih.gov/pubmed/29796416
http://dx.doi.org/10.1097/TXD.0000000000000779
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