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Department of Physiology, National Cheng-Kung University Medical College, Tainan, Taiwan 701, Republic of China
To study the effects of flow on in situ
endothelial intracellular calcium concentration
([Ca2+]i) signaling, rat aortic rings were
loaded with fura 2, mounted on a tissue flow chamber, and divided into
control and flow-pretreated groups. The latter was perfused with buffer
at a shear stress of 50 dyns/cm2 for 1 h. Endothelial
[Ca2+]i responses to ACh or shear stresses
were determined by ratio image analysis. Moreover, ACh-induced
[Ca2+]i elevation responses were measured in
a calcium-free buffer, or in the presence of SKF-96365, to elucidate
the role of calcium influx in the flow effects. Our results showed that
1) ACh increased endothelial
[Ca2+]i in a dose-dependent manner, and these
responses were incremented by flow-pretreatment; 2) the
differences in ACh-induced [Ca2+]i elevation
between control and flow-pretreated groups were abolished by SKF-96365
or by Ca2+-free buffer; and 3) in the presence
of 10
5 M ATP, shear stress induced dose-dependent
[Ca2+]i elevation responses that were not
altered by flow-pretreatment. In conclusion, flow-pretreatment augments
the ACh-induced endothelial calcium influx in rat aortas ex vivo.
flow pretreatment; calcium image; acetylcholine; shear stress; endothelial cells
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