We previously have shown in mice and rats, enhanced leukocyte recruitment to airway post-capillary venules after excessive distention imposed by the application of positive end-expiratory pressure. Since p-selectin was shown to be essential for this outcome, we sought to establish an in vitro endothelial cell model and determine the mechanisms whereby mechanical distension alters adhesion molecule expression. P-selectin surface expression on mouse jugular vein endothelial cells exposed to cyclic stretch (5% or 20% elongation for 5 min; Flexercell) were compared to static cells. The larger, pathophysiologic regimen of cyclic stretch showed a 54% increase in p-selectin expression after stretch compared to static cells. This response was attenuated but confirmed in tracheal venular endothelium (29% increase). We questioned whether these changes were dependent on increases in intracellular Ca²⁺ through voltage-gated Ca²⁺ channels. The stretch-induced increase in p-selectin expression was substantially decreased by pretreatment with the T-type Ca²⁺ channel inhibitor mibefradil (76% inhibition). Furthermore, when the Ca_v3.1 T-type Ca²⁺ channel expression was decreased in both endothelial cell subtypes with specific siRNA, the distension-induced expression of p-selectin decreased to levels less than that observed in static cells. We conclude that p-selectin expression on systemic venular endothelial cells contributes to a pro-inflammatory phenotype after mechanical stretch and can be selectively modulated by voltage-gated calcium channel inhibition.