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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2018-05-01
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doaj-7a3620ec0b624d3399e347960721775c2020-11-24T21:58:21ZengWolters KluwerTransplantation Direct2373-87312018-05-0145e34510.1097/TXD.0000000000000779201805000-0005Endothelial Dysfunction in Steatotic Human Donor Livers: A Pilot Study of the Underlying Mechanism During Subnormothermic Machine PerfusionIrene Beijert, MD0Safak Mert, PhD1Viola Huang, MD2Negin Karimian, MD3Sharon Geerts, BSc4Ehab O.A. Hafiz, MD5James F. Markmann, MD6Heidi Yeh, MD7Robert J. Porte, MD, PhD8Korkut Uygun, PhD91 Center for Engineering in Medicine, Department of Surgery, Massachusetts General Hospital, Harvard Medical School, Boston, MA.1 Center for Engineering in Medicine, Department of Surgery, Massachusetts General Hospital, Harvard Medical School, Boston, MA.1 Center for Engineering in Medicine, Department of Surgery, Massachusetts General Hospital, Harvard Medical School, Boston, MA.1 Center for Engineering in Medicine, Department of Surgery, Massachusetts General Hospital, Harvard Medical School, Boston, MA.1 Center for Engineering in Medicine, Department of Surgery, Massachusetts General Hospital, Harvard Medical School, Boston, MA.1 Center for Engineering in Medicine, Department of Surgery, Massachusetts General Hospital, Harvard Medical School, Boston, MA.4 Transplant Center, Department of Surgery, Massachusetts General Hospital, Boston, MA.4 Transplant Center, Department of Surgery, Massachusetts General Hospital, Boston, MA.2 Section of Hepatobiliary Surgery and Liver Transplantation, Department of Surgery, University Medical Center Groningen, University of Groningen, Groningen, The Netherlands.1 Center for Engineering in Medicine, Department of Surgery, Massachusetts General Hospital, Harvard Medical School, Boston, MA.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.http://journals.lww.com/transplantationdirect/fulltext/10.1097/TXD.0000000000000779 |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Irene Beijert, MD Safak Mert, PhD Viola Huang, MD Negin Karimian, MD Sharon Geerts, BSc Ehab O.A. Hafiz, MD James F. Markmann, MD Heidi Yeh, MD Robert J. Porte, MD, PhD Korkut Uygun, PhD |
spellingShingle |
Irene Beijert, MD Safak Mert, PhD Viola Huang, MD Negin Karimian, MD Sharon Geerts, BSc Ehab O.A. Hafiz, MD James F. Markmann, MD Heidi Yeh, MD Robert J. Porte, MD, PhD Korkut Uygun, PhD Endothelial Dysfunction in Steatotic Human Donor Livers: A Pilot Study of the Underlying Mechanism During Subnormothermic Machine Perfusion Transplantation Direct |
author_facet |
Irene Beijert, MD Safak Mert, PhD Viola Huang, MD Negin Karimian, MD Sharon Geerts, BSc Ehab O.A. Hafiz, MD James F. Markmann, MD Heidi Yeh, MD Robert J. Porte, MD, PhD Korkut Uygun, PhD |
author_sort |
Irene Beijert, MD |
title |
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_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_sort |
endothelial dysfunction in steatotic human donor livers: a pilot study of the underlying mechanism during subnormothermic machine perfusion |
publisher |
Wolters Kluwer |
series |
Transplantation Direct |
issn |
2373-8731 |
publishDate |
2018-05-01 |
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. |
url |
http://journals.lww.com/transplantationdirect/fulltext/10.1097/TXD.0000000000000779 |
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