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Effect of Irradiation on Gene Expression of Rat Liver Adhesion Molecules

In Vivo and In Vitro Studies

Einfluss ionisierender Strahlung auf die Expression von Adhäsionsmolekülen in der Leber. In-vivo- und In-vitro-Studien

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Background and Purpose:

Migration of leukocytes into tissue is a key element of innate and adaptive immunity. An animal study showed that liver irradiation, in spite of induction of chemokine gene expression, does not lead to recruitment of leukocytes into the parenchyma. The aim of this study was to analyze gene expression of adhesion molecules, which mediate leukocyte recruitment into organs, in irradiated rat liver in vivo and rat hepatocytes in vitro.

Material and Methods:

Rat livers in vivo were irradiated selectively at 25 Gy. Isolated hepatocytes in vitro were irradiated at 8 Gy. RNA extracted within 48 h after irradiation in vivo and in vitro was analyzed by real-time PCR (polymerase chain reaction) and Northern blot. Adhesion molecule concentration in serum was measured by ELISA (enzyme-linked immunosorbent assay). Cryostat sections of livers were used for immunohistology.

Results:

Significant radiation-induced increase of ICAM-1 (intercellular adhesion molecule-1), VCAM-1 (vascular cell adhesion molecule-1), JAM-1 (junctional adhesion molecule-1), β1-integrin, β2-integrin, E-cadherin, and P-selectin gene expression could be detected in vivo, while PECAM-1 (platelet-endothelial cell adhesion molecule-1) gene expression remained unchanged. In vitro, β1-integrin, JAM-1, and ICAM-2 showed a radiation-induced increased expression, whereas the levels of P-selectin, ICAM-1, PECAM-1, VCAM-1, Madcam-1 (mucosal addressin cell adhesion molecule-1), β2-integrin, and E-cadherin were downregulated. However, incubation of irradiated hepatocytes with either tumor necrosis factor-(TNF-)α, interleukin-(IL-)1β, or IL-6 plus TNF-α led to an upregulation of P-selectin, ICAM-1 and VCAM-1.

Conclusion:

The findings suggest that liver irradiation modulates gene expression of the main adhesion molecules in vivo and in cytokine-activated hepatocytes, with the exception of PECAM-1. This may be one reason for the lack of inflammation in the irradiated rat liver.

Hintergrund und Ziel:

Tierexperimentelle Studien haben gezeigt, dass es in der Akutphase nach Einzeitbestrahlung der Leber mit 25 Gy im Gegensatz zu anderen toxischen Leberschädigungen trotz strahleninduzierter Expression von Chemokinen nicht zu einer inflammatorischen Reaktion mit Einwanderung von Entzündungszellen kommt. Ziel der vorliegenden experimentellen Studie war die Messung der Auswirkungen ionisierender Strahlung auf die Expression der wichtigsten Adhäsionsmoleküle nach Leberbestrahlung in vivo und Bestrahlung von Hepatozyten in vitro im etablierten Modell.

Material und Methodik:

Innerhalb von 48 h nach selektiver Leberbestrahlung in vivo (25 Gy) sowie Bestrahlung von Hepatozyten in vitro (8 Gy) wurde RNA extrahiert und mittels Real-Time-PCR (Polymerase-Kettenreaktion) und Northern-Blot analysiert. Neben alleinig bestrahlten Hepatozyten wurden dabei in vitro auch Zellen untersucht, die zusätzlich zur Bestrahlung mit Tumor-Nekrose-Faktor-(TNF-)α, Interleukin-(IL-)1β oder einer Kombination aus IL-6/TNF-α inkubiert wurden. Adhäsionsmolekülkonzentrationen im Serum wurden mittels ELISA („enzyme-linked immunosorbent assay“) gemessen, Lebergewebe auch mittels Immunhistochemie untersucht.

Ergebnisse:

ICAM-1 („intercellular adhesion molecule-1“), VCAM-1 („vascular cell adhesion molecule-1“), JAM-1, („junctional adhesion molecule-1“), β1-Integrin, β2-Integrin, E-Cadherin und P-Selectin waren in vivo nach Bestrahlung vermehrt exprimiert, die PECAM-1-Expression („platelet-endothelial cell adhesion molecule-1“) blieb jedoch unverändert. In vitro kam es zu einer vermehrten Expression von β1-Integrin, JAM-1 und ICAM-2 und einer verminderten Expression von P-Selectin, ICAM-1, PECAM-1, VCAM-1, Madcam-1 („mucosal addressin cell adhesion molecule 1“), β2-Integrin und E-Cadherin. Nach zusätzlicher Inkubation der Hepatozyten mit TNF-α, IL-1β oder IL-6 und TNF-α kam es auch in vitro zu einer vermehrten Expression von P-Selectin, ICAM-1 und VCAM-1.

Schlussfolgerung:

Leberbestrahlung führt zu einer vermehrten Expression der wichtigsten Adhäsionsmoleküle in vivo und in durch Zytokine aktivierten Hepatozyten. Die PECAM-1-Expression wird allerdings nicht beeinflusst. Dies könnte einer der Gründe für die fehlende Inflammation in diesem Modell sein.

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Authors and Affiliations

  1. Department of Gastroenterology and Endocrinology, Göttingen University, Göttingen, Germany

    Federico Moriconi, Ihtzaz Malik, Ghayyor Ahmad, Joszef Dudas & Giuliano Ramadori

  2. Department of Radiotherapy, Göttingen University, Göttingen, Germany

    Margret Rave-Fränk, Hilke Vorwerk, Andrea Hille, Clemens Friedrich Hess &  Hans Christiansen

  3. Klinik und Poliklinik für Strahlentherapie und Radioonkologie, Universitätsmedizin Göttingen, Robert-Koch-Straße 40, 37075, Göttingen, Germany

    Hans Christiansen

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  1. Federico Moriconi
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  2. Ihtzaz Malik
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  3. Ghayyor Ahmad
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  4. Joszef Dudas
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  5. Margret Rave-Fränk
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  9. Giuliano Ramadori
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Correspondence to Hans Christiansen.

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Moriconi, F., Malik, I., Ahmad, G. et al. Effect of Irradiation on Gene Expression of Rat Liver Adhesion Molecules. Strahlenther Onkol 185, 460–468 (2009). https://doi.org/10.1007/s00066-009-1964-1

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  • DOI: https://doi.org/10.1007/s00066-009-1964-1

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