Direct, real-time measurement of shear stress-induced nitric oxide produced from endothelial cells in vitro.
Direct, real-time measurement of shear stress-induced nitric oxide produced from endothelial cells in vitro.
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DOI：
10.1016/j.niox.2010.08.003
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发表时间：
2010-12-15
期刊：
影响因子：
--
通讯作者：
Barbee KA
中科院分区：
文献类型：
--
作者：
Andrews AM;Jaron D;Buerk DG;Kirby PL;Barbee KA
内皮细胞产生的一氧化氮（NO）参与血管张力的调节。NO产生或可用性降低与高胆固醇血症和高血压中的内皮功能障碍有关。剪切应力诱导的NO释放是一个公认的现象，但这种反应的细胞机制尚未完全理解。实验的局限性阻碍了直接，实时测量NO流动条件下。我们已经克服了这些挑战与一个新的设计，平行板流动室。该室由两个隔室组成，由Transwell®膜隔开，该膜将位于上隔室中的NO记录电极与流动效应隔离。内皮细胞生长在膜的底部，膜插入与上板齐平的腔室中。我们证明了第一次直接实时NO测量从内皮细胞的剪切应力的控制变化。剪切应力从0.1达因/厘米2到6，10或20达因/厘米2的阶跃变化引起NO的瞬时减少，然后增加到一个新的稳态。NO运输的分析表明，最初的减少是由于对流的去除率随着流量的增加而增加。此外，NO浓度接近新的稳态的速率与时间依赖性细胞反应有关，而不是测量配置的运输限制。我们的设计提供了一种方法，用于研究动力学的信号机制连接NO的生产与剪切应力以及病理条件，涉及NO的生产或可用性的变化。
Nitric oxide (NO) produced by the endothelium is involved in the regulation of vascular tone. Decreased NO production or availability has been linked to endothelial dysfunction in hypercholesterolemia and hypertension. Shear stress-induced NO release is a well-established phenomenon, yet the cellular mechanisms of this response are not completely understood. Experimental limitations have hindered direct, real-time measurements of NO under flow conditions. We have overcome these challenges with a new design for a parallel-plate flow chamber. The chamber consists of two compartments, separated by a Transwell® membrane, which isolates a NO recording electrode located in the upper compartment from flow effects. Endothelial cells are grown on the bottom of the membrane, which is inserted into the chamber flush with the upper plate. We demonstrate for the first time direct real-time NO measurements from endothelial cells with controlled variations in shear stress. Step changes in shear stress from 0.1 dyn/cm2 to 6, 10 or 20 dyn/cm2 elicited a transient decrease in NO followed by an increase to a new steady state. An analysis of NO transport suggests that the initial decrease is due to the increased removal rate by convection as flow increases. Furthermore, the rate at which the NO concentration approaches the new steady state is related to the time-dependent cellular response rather than transport limitations of the measurement configuration. Our design offers a method for studying the kinetics of the signaling mechanisms linking NO production with shear stress as well as pathological conditions involving changes in NO production or availability.
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影响因子：
3.2
作者：
Mochizuki, S;Goto, M;Kajiya, F
通讯作者：
Kajiya, F
影响因子：
37.8
作者：
Tsao, PS;Buitrago, R;Cooke, JP
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Cooke, JP
影响因子：
3.3
作者：
Berkels, R;Purol-Schnabel, S;Roesen, R
通讯作者：
Roesen, R
DOI：
10.1073/pnas.88.11.4651
发表时间：
1991-06-01
影响因子：
11.1
作者：
KUBES, P;SUZUKI, M;GRANGER, DN
通讯作者：
GRANGER, DN
影响因子：
4.8
作者：
Espey, MG;Miranda, KM;Wink, DA
通讯作者：
Wink, DA