ELECTROPHARMACOLOGYCAL ANALYSIS OF INTRACELLULAR SIGNALTRANSDUCTIONS OF VASCULAR ENDOTHERIAL CELLS

血管内皮细胞胞内信号传导的电药理学分析

基本信息

批准号：
06670098
负责人：
IIJIMA Toshihiko
金额：
$ 1.34万
依托单位：
AKITA UNIVERSITY
依托单位国家：
日本
项目类别：
Grant-in-Aid for General Scientific Research (C)
财政年份：
1994
资助国家：
日本
起止时间：
1994 至 1995
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-06670098/
关键词：
aortic endotherial cell intracelullaru Ca^<2+> concentration Thapsigargin Cyclopiazonic acid endoplasmic Ca^<2+> ATPase Ca^<2+> entry Calcium influx factor (CIF)Intracellular Ca^<2+> store 小胞体Ca^<2+> -ATPase 小胞体Ca^<2+>-ATPase

项目摘要

In cultured vascular endothelial cells, histamine and ATP at high concentrations (>10 muM) produce an initial transient followed by sustained elevation in intracellular free Ca^<2+> concentration ([Ca^<2+>]_i). In the present experiments, mechanisms responsible for the latter sustained elevation were studied in cultured human aortic endothelial cells loaded with the fluorescent Ca^<2+> indicator fura-2. The latter phase was eliminated by removal of extracellular Ca^<2+>, and was reversibly abolished by reduction of extracellular Cl^- concentration to 40 mM or by the Cl^- channel blocker N-phenylanthranilic acid (NPA,1 mM). Effects of thapsigargin and cyclopiazonic acid (CPA), specific inhibitors of endoplasmic reticulum Ca^<2+>-ATPase, on [Ca^<2+>]_i were also examined. Thapsigargin (1-1000 nM) and CPA (0.1-100 muM) produced a biphasic change in [Ca^<2+>]_i. The slow declining phase was eliminated by removal of extracellular Ca^<2+> and was prevented by the low Cl^- condition and NPA.These results suggest that the sustained elevation of [Ca^<2+>]_i in response to histamine, ATP,thapsigargin and CPA is produced by the Cl^--sensitive entry of extracellular Ca^<2+> activated by the rise in [Ca^<2+>]_i and/or by the depletion of intracellular Ca^<2+> stores in cultured human aortic endothelial cells.

在培养的血管内皮细胞中，组胺和ATP高浓度（> 10 mum）产生初始瞬态，然后在细胞内游离Ca^<2+>浓度（[Ca^<2+>] _ I）中持续升高。在目前的实验中，在培养的人主动脉内皮细胞中研究了负责后者持续升高的机制，并带有荧光Ca^<2+>指示剂Fura-2。通过去除细胞外Ca^<2+>来消除后期，并通过将细胞外Cl^ - 浓度降低至40 mm或通过Cl^ - 通道阻滞剂N-苯基氨基酸（NPA，NPA，1 mm）而可逆地废除。还检查了[Ca^<2+>] i的thapsigargin和环皮二唑酸（CPA）的影响，内质网ca^<2+> - ATPase的特定抑制剂。 Thapsigargin（1-1000 nm）和CPA（0.1-100 MUM）在[Ca^<2+>] _ I中产生了双相变化。通过去除细胞外Ca^<2+>消除了缓慢的阶段，并被低Cl^ - 条件和NPA所阻止。这些结果表明，响应组胺，ATP，Thapsigargin和Cpa的cl^siver ca-ca-ca-ca riped ca i>激活了[Ca^<2+>] _ I的持续升高[Ca^<2+>] _ i的持续升高，[ca^<2+>] _ i。和/或通过培养的人主动脉内皮细胞中细胞内Ca^<2+>存储的耗竭。