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Am J Physiol Heart Circ Physiol 289: H2193-H2202, 2005. First published July 1, 2005; doi:10.1152/ajpheart.00355.2005
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Clusterin: a protective mediator for ischemic cardiomyocytes?

P. A. J. Krijnen,1,5 S. A. G. M. Cillessen,1 R. Manoe,6 A. Muller,3,5 C. A. Visser,2,5 C. J. L. M. Meijer,1 R. J. P. Musters,3,5 C. E. Hack,4,5,6 L. A. Aarden,6 and H. W. M. Niessen1,5

Departments of 1Pathology, 2Cardiology, 3Physiology, and 4Clinical Chemistry, Vrije Universiteit Medical Center, Amsterdam, The Netherlands; 5Institute for Cardiovascular Research, Vrije Universiteit, Amsterdam, The Netherlands; and 6Sanquin Research, Central Laboratory for Blood Transfusion, Department of Immunopathology and Laboratory of Experimental and Clinical Immunology, Academic Medical Center, University of Amsterdam, Amsterdam, The Netherlands

Submitted 11 April 2005 ; accepted in final form 29 June 2005

We examined the relationship between clusterin and activated complement in human heart infarction and evaluated the effect of this protein on ischemic rat neonatal cardiomyoblasts (H9c2) and isolated adult ventricular rat cardiomyocytes as in vitro models of acute myocardial infarction. Clusterin protects cells by inhibiting complement and colocalizes with complement on jeopardized human cardiomyocytes after infarction. The distribution of clusterin and complement factor C3d was evaluated in the infarcted human heart. We also analyzed the protein expression of clusterin in ischemic H9c2 cells. The binding of endogenous and purified human clusterin on H9c2 cells was analyzed by flow cytometry. Furthermore, the effect of clusterin on the viability of ischemically challenged H9c2 cells and isolated adult ventricular rat cardiomyocytes was analyzed. In human myocardial infarcts, clusterin was found on scattered, morphologically viable cardiomyocytes within the infarcted area that were negative for complement. In H9c2 cells, clusterin was rapidly expressed after ischemia. Its expression was reduced after reperfusion. Clusterin bound to single annexin V-positive or annexin V and propidium iodide-positive H9c2 cells. Clusterin inhibited ischemia-induced death in H9c2 cells as well as in isolated adult ventricular rat cardiomyocytes in the absence of complement. We conclude that ischemia induces the upregulation of clusterin in ischemically challenged, but viable, cardiomyocytes. Our data suggest that clusterin protects cardiomyocytes against ischemic cell death via a complement-independent pathway.

ischemia; acute myocardial infarction; membrane flip-flop


Address for reprint requests and other correspondence: P. A. J. Krijnen, VU Univ. Medical Center, Dept. of Pathology, De Boelelaan 1117, 1007 MB Amsterdam, The Netherlands (e-mail: paj.krijnen{at}vumc.nl)




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