Matrix metalloproteinases-2 and -9 (MMP-2/9) are involved in the degradation of extracellular matrix, and their inhibition is discussed as a promising strategy against hepatic ischemia-reperfusion (I/R) injury. Here, we analyzed the role of MMP-2 and -9 for leukocyte migration in sham-operated mice and in mice after I/R treated either with a MMP-2/9 inhibitor or vehicle. We show that I/R-induced MMP-9 activation was abolished by the MMP-2/9 inhibitor, whereas MMP-2 activity was not detectable in all groups. As demonstrated by intravital microscopy, MMP-9 inhibition attenuated postischemic rolling and adherence of total leukocytes in venules, CD4+ T cell accumulation in sinusoids, and neutrophil transmigration. These effects were associated with a reduction of CD62P expression and plasma TNF-alpha levels. As shown by RLOT microscopy, motility of interstitially migrating leukocytes was significantly affected upon MMP-9 blockade. Postischemic sinusoidal perfusion failure, apoptosis, and ALT activity were only slightly reduced after MMP-9 inhibition, whereas AST activity and mortality were significantly lower. In conclusion, MMP-9 is involved in the early cascades of leukocyte recruitment and migration during hepatic I/R probably via TNF-alpha-mediated CD62P translocation, and is required for motility of interstitially migrating leukocytes. MMP-9 blockade weakly protects from early I/R damage, but improves survival.