Antiapoptotic Effect of Tacrolimus on Cytokine-Challenged Human Islets
Our goal was to investigate whether previously related antiapoptotic and anti-inflammatory effects of tacrolimus could be useful in protecting human islets cultured in the presence of several proinflammatory mediators. Human islets obtained from cadaveric donors after intraductal infusion with colla...
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2009-11-01
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Series: | Cell Transplantation |
Online Access: | https://doi.org/10.3727/096368909X12483162197240 |
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doaj-6606153ab0ba4d7091a286fc066807942020-11-25T03:24:48ZengSAGE PublishingCell Transplantation0963-68971555-38922009-11-011810.3727/096368909X12483162197240Antiapoptotic Effect of Tacrolimus on Cytokine-Challenged Human IsletsJose M. Balibrea Del Castillo M.D., Ph.D.0M. Cruz García-Martín1Javier Arias-Díaz2Elena Giné3Elena Vara4Jose L. Balibrea Cantero5 Department of Surgery, Universidad Complutense, Madrid, Spain Department of Biochemistry and Molecular Biology, Universidad Complutense, Madrid, Spain Department of Surgery, Universidad Complutense, Madrid, Spain Department of Biochemistry and Molecular Biology, Universidad Complutense, Madrid, Spain Department of Biochemistry and Molecular Biology, Universidad Complutense, Madrid, Spain Department of Surgery, Universidad Complutense, Madrid, SpainOur goal was to investigate whether previously related antiapoptotic and anti-inflammatory effects of tacrolimus could be useful in protecting human islets cultured in the presence of several proinflammatory mediators. Human islets obtained from cadaveric donors after intraductal infusion with collagenase, mechanical digestion, and continuous Ficoll gradient purification were cultured in RPMI-1640 medium for 24 h. Escherichia coli lipopolysaccharide (10 μg/ml) or interleukin-1 (50 UI/ml)+γ-IF (1000 UI/ml) and low-dose tacroliumus (5 ng/ml) were added. Homogenized samples (300 IE) from five different donors where assigned to four different experimental groups (control, treatment, cytokines, and cytokines + treatment). To evaluate islet damage and apoptotic response, nucleosome content, Bcl-2 protein levels, caspase-3, -8, and -9 levels, and insulin concentration were measured. Also, TNF-α and IL-6 levels where assessed as indicators of the inflammatory response. All proapoptotic markers, TNF-α, and IL-6 levels were augmented after both LPS and cytokine stimulation. Tacrolimus reduced significantly all of them and restored baseline values of nucleosome and caspase-9 in both experiments and Bcl-2 and caspase-3 when IL-1 + γ-IF was added. Twenty-four-hour insulin concentration diminished when LPS or IL-1 + γ-IF were present. Tacrolimus treatment restored insulin levels in both experiments. These results suggest that in vitro apoptotic events and media insulin concentration decrease after proinflammatory stimulation can be reverted using low-dose tacrolimus.https://doi.org/10.3727/096368909X12483162197240 |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Jose M. Balibrea Del Castillo M.D., Ph.D. M. Cruz García-Martín Javier Arias-Díaz Elena Giné Elena Vara Jose L. Balibrea Cantero |
spellingShingle |
Jose M. Balibrea Del Castillo M.D., Ph.D. M. Cruz García-Martín Javier Arias-Díaz Elena Giné Elena Vara Jose L. Balibrea Cantero Antiapoptotic Effect of Tacrolimus on Cytokine-Challenged Human Islets Cell Transplantation |
author_facet |
Jose M. Balibrea Del Castillo M.D., Ph.D. M. Cruz García-Martín Javier Arias-Díaz Elena Giné Elena Vara Jose L. Balibrea Cantero |
author_sort |
Jose M. Balibrea Del Castillo M.D., Ph.D. |
title |
Antiapoptotic Effect of Tacrolimus on Cytokine-Challenged Human Islets |
title_short |
Antiapoptotic Effect of Tacrolimus on Cytokine-Challenged Human Islets |
title_full |
Antiapoptotic Effect of Tacrolimus on Cytokine-Challenged Human Islets |
title_fullStr |
Antiapoptotic Effect of Tacrolimus on Cytokine-Challenged Human Islets |
title_full_unstemmed |
Antiapoptotic Effect of Tacrolimus on Cytokine-Challenged Human Islets |
title_sort |
antiapoptotic effect of tacrolimus on cytokine-challenged human islets |
publisher |
SAGE Publishing |
series |
Cell Transplantation |
issn |
0963-6897 1555-3892 |
publishDate |
2009-11-01 |
description |
Our goal was to investigate whether previously related antiapoptotic and anti-inflammatory effects of tacrolimus could be useful in protecting human islets cultured in the presence of several proinflammatory mediators. Human islets obtained from cadaveric donors after intraductal infusion with collagenase, mechanical digestion, and continuous Ficoll gradient purification were cultured in RPMI-1640 medium for 24 h. Escherichia coli lipopolysaccharide (10 μg/ml) or interleukin-1 (50 UI/ml)+γ-IF (1000 UI/ml) and low-dose tacroliumus (5 ng/ml) were added. Homogenized samples (300 IE) from five different donors where assigned to four different experimental groups (control, treatment, cytokines, and cytokines + treatment). To evaluate islet damage and apoptotic response, nucleosome content, Bcl-2 protein levels, caspase-3, -8, and -9 levels, and insulin concentration were measured. Also, TNF-α and IL-6 levels where assessed as indicators of the inflammatory response. All proapoptotic markers, TNF-α, and IL-6 levels were augmented after both LPS and cytokine stimulation. Tacrolimus reduced significantly all of them and restored baseline values of nucleosome and caspase-9 in both experiments and Bcl-2 and caspase-3 when IL-1 + γ-IF was added. Twenty-four-hour insulin concentration diminished when LPS or IL-1 + γ-IF were present. Tacrolimus treatment restored insulin levels in both experiments. These results suggest that in vitro apoptotic events and media insulin concentration decrease after proinflammatory stimulation can be reverted using low-dose tacrolimus. |
url |
https://doi.org/10.3727/096368909X12483162197240 |
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