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Vascular endothelial cells (VEC) provide an essential protective barrier between the vascular system and underlying tissues. Using VEC barrier models of human coronary artery cells and human and rat brain microvascular endothelial cells, we investigated the mechanism by which morphine affects lipopolysaccharide (LPS)-induced VEC permeability. We demonstrated that co-administration of morphine and LPS induced greater VEC apoptosis and permeability than morphine or LPS alone. The extent of induced apoptosis appeared to be cell-type dependent. Furthermore, RT-PCR analysis revealed that morphine and LPS up-regulated Fas expression. These data suggest potential crosstalk between the signaling pathways that mediate morphine- and LPS-triggered apoptosis in brain VEC.