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Rho-kinase as a novel gene therapeutic target in treatment of cold ischemia/reperfusion-induced acute lethal liver injury: effect on hepatocellular NADPH oxidase system

Gene Therapy volume 14, pages 1425–1433 (2007)Cite this article

Abstract

In the transplant surgery, reactive oxygen species (ROS) from the reperfused tissue cause ischemia-reperfusion injury, resulting in the primary graft failure. We have recently reported that Rho-kinase, an effecter of the small GTPase Rho, plays an important role in the ROS production in the hyperacute phase of reperfusion; however, the sources and mechanisms of the ROS production remain to be elucidated. The aim of this study was to investigate the source of ROS production with a special reference to Rho-kinase to develop a new strategy against ischemia-reperfusion injury. In an in vivo rat model of liver transplantation, Kupffer cells in the graft were depleted using liposome-encapsulated dichloromethylene diphosphonate to examine the source of ROS production. The effect of adenoviral-mediated overexpression of a dominant-negative Rho-kinase (AdDNRhoK) in hepatocytes in the graft was also examined. Kupffer cells were not involved in the ROS production, whereas the AdDNRhoK transfection to hepatocytes significantly suppressed the ROS production. Furthermore, the ROS production was dose-dependently inhibited by apocynin, an NADPH oxidase inhibitor. Expression of DNRhoK also suppressed the release of pro-inflammatory cytokines, and ameliorated the lethal liver injury with a significant prolongation of the survival. These results suggest that the Rho-kinase-mediated pathway plays a crucial role in the ROS production through NADPH oxidase in hepatocytes during the hyperacute phase of reperfusion in vivo. Thus, Rho-kinase in hepatocytes may be a new therapeutic target for the prevention of primary graft failure in liver transplantation.

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Abbreviations

ALT:

alanine aminotransferase

AP-1:

activator protein-1

AST:

aspartate aminotransferase

Cl2MDP:

dichloromethylene diphosphonate

COX:

cycloxygenase

IL-1β:

interleukin-1β

NF-κβ:

nuclear factor-κβ

ROS:

reactive oxygen species

TNF-α:

tumor necrosis factor-α

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Acknowledgements

We thank T Hattori, M Higashi, S Masuda and Y Kubota, Kyushu University Graduate School of Medical Sciences, for excellent technical assistance. This study was supported in part by the grant-in-aid (no. 13357011) and the grant for the 21st Century COE Program from the Japanese Ministry of Education, Culture, Sports, Science and Technology, Tokyo, Japan, and the Program for Promotion of Fundamental Studies in Health Sciences of the Organization for Pharmaceutical Safety and Research of Japan.

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Authors and Affiliations

  1. Department of Surgery and Science, Graduate School of Medical Sciences, Kyushu University, Fukuoka, Japan

    S Shiotani, A Taketomi, Y Soejima, K Hashimoto & Y Maehara

  2. Department of Digestive and Pediatric Surgery, Institute of Health Biosciences, The University of Tokushima Graduate School, Tokushima, Japan

    M Shimada & T Yoshizumi

  3. Department of Cardiovascular Medicine, Tohoku University, Graduate School of Medicine, Sendai, Japan

    H Shimokawa

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  1. S Shiotani
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Correspondence to S Shiotani.

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Shiotani, S., Shimada, M., Taketomi, A. et al. Rho-kinase as a novel gene therapeutic target in treatment of cold ischemia/reperfusion-induced acute lethal liver injury: effect on hepatocellular NADPH oxidase system. Gene Ther 14, 1425–1433 (2007). https://doi.org/10.1038/sj.gt.3303000

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