Development of the optical system for continuous and multi-factorial monitoring of the intracellular signaling network in endothelial and smooth muscle cells in vascular strips.

开发光学系统，用于连续和多因素监测血管条内皮细胞和平滑肌细胞的细胞内信号网络。

• 批准号: 10557072
• 负责人: KANAIDE Hideo
• 金额: $ 8.32万
• 依托单位: KYUSHU UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B).
• 财政年份: 1998
• 资助国家: 日本
• 起止时间: 1998 至 2000
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-10557072/
• 关键词: intracellular signaling network; endothelial cells; vascular sooth muscle cells; fura-2; BCECF; optical fibers; regulation of vascular tonus; タプシガルギン; 容量依存性性カルシウム流入

The purpose of this study is to develop optical systems for continuous and multi-factorial monitoring of the intracellular signaling network in endothelial and smooth muscle cells in vascular strips. Using newly developed systems, we investigated the mechanisms underlying the regulation of the vascular tonus. Following results were obtained in this study : (1) We have developed a system to monitor changes in [Ca^<2+>]i and tension of the vascular strips simultaneously. This system was reported as a chapter "Measurement of [Ca^<2+>]i in smooth muscle strips using front-surface fluorimetry" in the book "Calcium signaling protocols" edited by David G.Lambert, which is the Vol.114 of series of "Methods in Molecular Biology" (Humana Press Inc.1999). (2) Using front-surface fluorimetry of fura-2, we investigated the mechanism underlying the three phasic endothelium-dependent responses induced by thapsigargin during the phenirephrine-induced contraction in porcine renal small arterial strips. … More It was found that the initial transient relaxation accompanied by a reduction of smooth muscle [Ca^<2+>]i was due to both the release of endothelium-derived hyperpolarizing factor(EDHF) and nitric oxide(NO).. The subsequent transient contraction accompanied by an increase in [Ca^<2+>]i was due a release of thromboxane A_2 from the endothelium. The final sustained relaxation, which was not accompanied by the changes in[Ca^<2+>]i was due to release of NO from the endothelium.(3)We have developed a new system to monitor [Ca^<2+>]i and (pH)i of vascular smooth muscle cells simultaneously. (4) Using this system, we investigated the mechanism underlying an increase in the tension development induced by the alkalization of smooth muscle cells. The application of NH_4Cl induced an alkalization which is accompanied by an increase in[Ca^<2+>]i in rat aortic smooth muscle cells in primary culture. It was found that alkalization increases Ca^<2+> entry through the SKF96365-sensitive Ca^<2+> channels. The capacitative entry of Ca^<2+> through cell membrane might increase [Ca^<2+>]i, and thus, increases tension development during alkalization. Less

本研究的目的是开发光学系统，用于连续和多因素监测血管条中内皮细胞和平滑肌细胞的细胞内信号网络。使用新开发的系统，我们研究了血管紧张性调节的机制。（1）建立了一套同时监测血管条[Ca^<2+>]i和张力变化的系统。该系统在大卫G.兰伯特编辑的“钙信号协议”一书中的“使用前表面荧光测定法测量平滑肌条中的[Ca^<2+>]i”一章中报道，该书是“分子生物学方法”系列的第114卷（Humana Press Inc.1999）。(2)利用Fura-2荧光探针研究了在苯肾上腺素诱导的猪肾小动脉条收缩过程中，毒胡萝卜素诱导的三个阶段的内皮依赖性反应的机制。 ...更多信息 结果发现，伴随着平滑肌[Ca^2+]i降低的初始瞬时舒张是由于内皮源性超极化因子（EDHF）和一氧化氮（NO）的释放。随后的短暂收缩伴随[Ca^<2+>]i的增加是由于内皮释放血栓素A_2所致。内皮细胞释放NO是最终持续舒张的原因，而不伴随[Ca^2+]i的变化。(3)We已经开发了一种新的系统来同时监测血管平滑肌细胞的[Ca^<2+>]i和（pH）i。(4)使用这个系统，我们调查的基础上增加的张力发展的平滑肌细胞的碱化诱导的机制。应用NH_4Cl诱导原代培养的大鼠主动脉平滑肌细胞碱性化，并伴有[Ca^<2+>]i增加。研究发现，碱化增加了Ca^2+通过SKF 96365敏感性Ca^2+通道的内流。Ca^<2+>通过细胞膜的容性进入可能增加[Ca^<2+>]i，从而增加碱化过程中张力的发展。少

项目成果

Ihara E, Hirano K, Nishimura J, Nawata H, Kanaide H: "Thapsigargin-induced endothelium-dependent triphasic regulation of vascular tone in the porcine renal artery."Br J Pharmacol. 128. 689-699 (1999)

Ihara E、Hirano K、Nishimura J、Nawata H、Kanaide H：“毒胡萝卜素诱导的猪肾动脉血管张力的内皮依赖性三相调节。”Br J Pharmacol。

Morisaki T, Matsunaga H, Beppu K, Ihara E, Hirano K, Kanaide H, Mori M, Katano M: "A combination of cyclosporin-A (CsA) and interferon-gamma (INF-γ)induces apoptosis in human gastric carcinoma cells."Anticancer Research. 20. 3363-3374 (2000)

Morisaki T、Matsunaga H、Beppu K、Ihara E、Hirano K、Kanaide H、Mori M、Katano M：“环孢素 A (CsA) 和干扰素-γ (INF-γ) 的组合可诱导人胃癌细胞凋亡.“抗癌研究。20. 3363-3374 (2000)

Matoba T: "Hydrogen peroxide is an endothelilum-derived hyperpolarizing factor in mice."J Clin Inv. 106. 1521-1530 (2000)

Matoba T：“过氧化氢是小鼠内皮衍生的超极化因子。”J Clin Inv。

Mizuno O: "Mechanism of endothelium-dependent relaxation induced by thrombin in the pig coronary artery."Eur J Pharmacol. 351. 67-77 (1998)

Mizuno O：“凝血酶在猪冠状动脉中诱导内皮依赖性舒张的机制。”Eur J Pharmacol。

Takeda K: "Peroxisome proliferator-activated receptor γ activators downregulate angiotensin II type 1 receptor in vascular smooth muscle cells."Circulation. 102. 1834-1839 (2000)

Takeda K：“过氧化物酶体增殖物激活受体 γ 激活剂下调血管平滑肌细胞中的血管紧张素 II 1 型受体。”循环。102。1834-1839 (2000)