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

• 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

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，高浓度(>10μM)的组胺和ATP产生最初的瞬时变化，随后细胞内游离Ca 2+ 浓度([Ca 2+ ]_i)持续升高。在本实验中，在负载有荧光Ca 2+ 指示剂fura-2的培养的人主动脉内皮细胞中研究了造成后者持续升高的机制。后一相通过去除细胞外Ca 2+ 来消除，并且通过将细胞外Cl 2- 浓度降低至40mM或通过Cl 2- 通道阻断剂N-苯基邻氨基苯甲酸(NPA，1mM)可逆地消除。还检测了毒胡萝卜素和环吡嗪酸(CPA)（内质网Ca^2+-ATP酶的特异性抑制剂）对[Ca^2+]_i的影响。毒胡萝卜素(1-1000nM)和CPA(0.1-100μM)产生[Ca^2+]_i的双相变化。缓慢下降期通过去除细胞外 Ca^<2+> 来消除，并通过低 Cl^- 条件和 NPA 来阻止。这些结果表明，[Ca^<2+>]_i 响应组胺而持续升高。 、 ATP、毒胡萝卜素和 CPA 是由 [Ca^<2+>]_i 的增加和/或细胞内 Ca^<2+> 的消耗所激活的 Cl^-敏感的细胞外 Ca^<2+> 进入而产生的。 Ca 2+ 储存在培养的人主动脉内皮细胞中。

项目成果

E.Hosoki&T.Iijima: "Chloride-sensitive Ca^<2+> entry by histamine and ATP in human aortic endothelial cells." Eur.J.Pharmacol.(Mol.Pharmacol.Sec.). 266. 213-218 (1994)

细木

E.Hosoki & T.Iijima: "Modulation of cytosolic Ca^<2+> concentration by thapsigargin and cyclopiazonic acid in human aortic endothelial cells." Eur. J. Pharmacol. (Mol. Pharmacol. Sec.). 288. 131‐137 (1995)

E.Hosoki 和 T.Iijima：“毒胡萝卜素和环吡嗪酸对人主动脉内皮细胞中胞质 Ca^2+ 浓度的调节”。Eur. J. Pharmacol. 288. 131‐。 137 (1995)

E.Hosoki & T.Iijima: "Modulation of cytosolic Ca^<2+> concentration by thapsigargin and cyclopiazonic acid in human aortic endothelial cells." Eur.J.Pharmacol.(Mol.Pharmacol.Sec.). 288. 131-137 (1995)

细木

E.Hosoki & T.Iijima: "Modulation of cytosolic Ca^<2+> concentration by thapsigargin and cyclopiazonic acid in human aortic endothelial cells" Eur.J.Pharmacol.(Mol.Pharmacol.Sec.). 288. 131-137 (1995)

细木

E.Hosoki&T.Iijima: "Modulation of cytosolic Ca^<2+> concentration by thapsigargin and cyclopiazonic acid in human aortic endothelial cells." Eur.J.Pharmacol.(Mol.Pharmacol.Sec.). 288. 131-137 (1995)

细木