Assessement of smooth muscle viability in the spastic cerebral arteries during chronic vasospasm

慢性血管痉挛期间痉挛脑动脉平滑肌活力的评估

• 批准号: 05671149
• 负责人: KIM Phyo
• 金额: $ 1.34万
• 依托单位: UNIVERSITY OF TOKYO
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (C)
• 财政年份: 1993
• 资助国家: 日本
• 起止时间: 1993 至 1994
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-05671149/
• 关键词: Cerebral vasospasm,; Subarachnoid hemorrhage,; Smooth muscle,; Permeability,; DNA-binding dye,; Apoptosis; TUNEL法; 膜透過性; 細胞内カルシウム

Alteration of sarcolemmal permeability was evaluated in the cerebral artery after subarachnoid hemorrhage (SAH). Significance of membrane dysfunction in the pathogenesis of chronic spasm and contribution of apoptosis were investigated in a canine model.Permeability of the smooth muscle cell (SMC) membrane was assessed by double staining with a hydrophilic (ethidium bromide : EB) and a lipophilic (Hoechst 33342 : H33342) DNA-binding dye. Quantitative observations were made with a UV-fluorescence microscope and a UV-laser confocal microscope. Occurrence of apoptosis was studied using electrophoresis and TUNEL method. In the normal arteries, nuclel of SMC were stained with H33342 but not with EB.In the spastic arteries obtained seven days after SAH,45(]SY.+-。[)19% of SMC in the inner layr of the tunica media were stained with EB.The incidence of EB-positive cells decreased in two to four weeks (13(]SY.+-。[)5.2% and 5.0(]SY.+-.[)2.1% respectively) in parallel with amelioration of spasm. Electron and light microscopic observations revealed increased density of SMC cytoplasm with widening of the extracellular space. Necrosis was not evident. Apoptosis was not detected by the two methods. These results demonstrated that an augmentation in sarcolemmal permeability takes place during the course of chronic vasospasm, and suggest its close correlation to pathogenesis.

评估蛛网膜下腔出血（SAH）后大脑动脉肌膜通透性的变化。在犬模型中研究了膜功能障碍在慢性痉挛发病机制中的重要性以及细胞凋亡的贡献。通过用亲水性（溴化乙锭：EB）和亲脂性（Hoechst 33342：H33342）DNA 结合染料双重染色来评估平滑肌细胞（SMC）膜的渗透性。使用紫外荧光显微镜和紫外激光共聚焦显微镜进行定量观察。使用电泳和TUNEL方法研究细胞凋亡的发生。正常动脉中，SMC细胞核被H33342染色，而EB不染色。SAH后7天获得的痉挛动脉中，45(]SY.+-。[)19%中膜内层SMC被EB染色。2~4周内EB阳性细胞发生率下降(13(]SY.+-。[)5.2%和 5.0(]SY.+-.[)2.1%)与痉挛的改善同时进行。电子显微镜和光学显微镜观察显示，随着细胞外空间的扩大，SMC 细胞质的密度增加。坏死不明显。两种方法均未检测到细胞凋亡。这些结果表明，在慢性血管痉挛过程中，肌膜通透性增加，并表明其与发病机制密切相关。

项目成果

Phyo Kim, Yuhei Yoshimoto, Hiroshi Nakaguchi, Takashi Mori, Akio Asai, Tomio Sasaki, Takaaki Kirino, and Yoshiaki Nonomura :"Increased Sarcolemmal Permeability in the Cerebral Artery during Chronic Spasm-An Assessment using DNA-binding Dyes and Detection

Phyo Kim、Yuhei Yoshimoto、Hiroshi Nakaguchi、Takashi Mori、Akio Asai、Tomio Sasaki、Takaaki Kirino 和 Yoshiaki Nonomura：“慢性痉挛期间脑动脉肌膜通透性增加 - 使用 DNA 结合染料和检测进行评估

Kim P, Yoshimoto Y, Nakaguchi H et al.: "Increased Sarcolemmal Permeability in the Cerebral Artery during Chronic Spasm" J Cereb Blood Flow Met. (submltted). (1998)

Kim P、Yoshimoto Y、Nakaguchi H 等人：“慢性痉挛期间脑动脉肌膜通透性增加”J Cereb Blood Flow Met。

Kim,P: "Loss of relaxations,metabolic farlure and increasad calcium permealility" J.Auton Nerv System. 49. S157-162 (1994)

Kim,P：“放松的丧失、代谢紊乱和钙渗透性增加”J.Auton 神经系统。

Kim,P et al: "dncreased calcium permealility of Sarcolemma during Vasospasm" J.Cereb.Blood Flow Met. (in press). (1995)

Kim,P 等人：“血管痉挛期间肌膜的钙渗透性降低”J.Cereb.Blood Flow Met。