Abstract
Endothelial cells (ECs) that line the inner surface of blood vessels are continuously exposed to shear stress induced by blood flow in human body, and shear stress affects the intracellular calcium ([Ca^<2+>]_i), which initiates cellular responses. Thus, we studied the intercellular communication of cultured ECs under shear stress. We used cultured bovine aortic cells and a parallel plate flow chamber for shear stress loading. We measured [Ca^<2+>]_i responses in cultured ECs by using a fluorescent microscope and calcium indicator (Calcium Green-1/AM), and measured [Ca^<2+>]_i, responses under shear stress in the presence of various inhibitors. The [Ca^<2+>]_i responses induced by shear stress in the presence of suramin (100μM), apyrase (10 units/ml), and FCCP (100μM) decreased to 40%, 25%, and 20%, respectively. However, the response in the presence of 18α-GA (gap junction inhibitor) did not change compared to no-treatment sample. Furthermore, after loading caged IP_3 into ECs, caged IP_3 was photoliberated in ECs. IP_3-triggered [Ca^<2+>]_i response propagated to adjacent cells downstream. These results have indicated the intercellular communication of ECs under shear stress is paracrine manner.
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