The Protective Function of Neutrophil Elastase Inhibitor in ... : Transplantation (original) (raw)

Basic and Experimental Research

The Protective Function of Neutrophil Elastase Inhibitor in Liver Ischemia/Reperfusion Injury

Uchida, Yoichiro; Freitas, Maria Cecilia S.; Zhao, Danyun; Busuttil, Ronald W.; Kupiec-Weglinski, Jerzy W.

Dumont-UCLA Transplant Center, Division of Liver and Pancreas Transplantation, Department of Surgery, David Geffen School of Medicine at UCLA, Los Angeles, CA.

This work was supported by NIH Grants RO1 DK062357, AI23847, AI42223, and The Dumont Research Foundation.

Address correspondence to: Jerzy W. Kupiec-Weglinski, M.D., Ph.D., Dumont-UCLA Transplant Center 77-120 CHS, 10833 Le Conte Avenue, Los Angeles, CA 90095.

E-mail: [email protected]

Y.U. participated in research design, performed all liver IRI experiments and most of follow-up molecular experimental testings, and wrote first draft of the manuscript; M.C.S.F. performed in immunohistology; D.Z. performed in vitro cell culture studies; R.W.B. participated in discussion and provided partial funding; J.W.K.-W. participated in research design, finalized the manuscript, and sponsored the project.

Received 10 November 2009. Revision requested 23 December 2009.

Accepted 12 January 2010.

Abstract

Background.

A neutrophil elastase (NE) inhibitor, Sivelestat, has been approved for the treatment of acute lung injury associated with systemic inflammation in humans. Some reports have also shown its protective effects in liver inflammatory states. We have recently documented the importance of NE in the pathophysiology of liver ischemia/reperfusion injury, a local Ag-independent inflammation response. This study was designed to explore putative cytoprotective functions of clinically available Sivelestat in liver ischemia/reperfusion injury.

Methods.

Partial warm ischemia was produced in the left and middle hepatic lobes of C57BL/6 mice for 90 min, followed by 6 or 24 hr of reperfusion. The mice were given Sivelestat (100 mg/kg, subcutaneous) at 10 min before ischemia, 10 min before reperfusion, and at 1 and 3 hr of reperfusion thereafter.

Results.

Sivelestat treatment significantly reduced serum alanine aminotransferase levels and NE activity, when compared with controls. Histological liver examination has revealed that unlike in controls, Sivelestat ameliorated the hepatocellular damage and decreased local neutrophil activity and infiltration. The expression of proinflammatory cytokines (tumor necrosis factor-α and interleukin-6), chemokines (CXCL-1, CXCL-2, and CXCL-10), and toll-like receptor 4 was significantly reduced in the treatment group, along with diminished apoptosis through caspase-3 pathway. Moreover, in vitro studies confirmed downregulation of proinflammatory cytokine and chemokine programs in mouse macrophage cell cultures, along with depression of innate toll-like receptor 4 signaling.

Conclusion.

Sivelestat-mediated NE inhibition may represent an effective therapeutic option in liver transplantation and other inflammation disease states.