探讨东茛萣素对血管平滑肌细胞内活性氧物质形成之影响.doc

探討東茛萣素對血管平滑肌細胞形成之影 Study of the effects of scopoletin on reactive oxygen species generation in vascular smooth muscle cells 研究生：楊彩微 Yang, Tsai-Wei 指導教授：黃卓治 Huang, Tzou-Chi 【摘要】 東茛萣素（6-methoxy-7-hydroxycoumarin, Scopoletin）一種酚類香豆素。根據研究顯示東茛萣素具有抗氧化、抗菌、抗發炎及保肝等功效外，也指出具有擴張血管達到降低血壓的作用。活性氧（Reactive oxygen species, ROS）在心血管疾病上扮演很重要角色，先前研究指出佛波醇-12-肉蔻酯-13-乙酯（phorbol 12-myristate 13-acetate, PMA）會刺激血管平滑肌細胞產生活性氧，PMA是強烈的非生理性刺激劑用於活化NADPH 氧化酶；NADPH 氧化酶為酵素複合體，其至少包含五個次單位：P47phox和ac、p22phox及 Nox1；當細胞受到刺激時，次單位p47蛋白會磷酸化，轉位至膜上與細胞內的次單位形成複合體。本研究目的探討東茛萣素對血管平滑肌細胞ROS形成之影響。由本次實驗結果可得知，Scopoletin具有濃度依賴性促使NO產生，而降低血管收縮劑（phenylephrine）所誘發血管段之收縮能力。東茛萣素具有降低PMA所誘發血管平滑肌細胞內ROS的產生和NADPH氧化酶活性，皆呈現出濃度及時間依賴性的抑制作用。利用西方墨點法分析，東茛萣素具有降低p47phox 蛋白表現量。此外，在共軛焦顯微鏡的結果顯示東茛萣素會降低PMA所誘導血管平滑肌細胞的p47phox轉位至膜上。這些結果說明東茛萣素能降低PMA所誘導ROS產生，是經由下游調節NADPH 氧化酶中的次單位P47phox蛋白表現量。、活性氧、血管平滑肌細胞、NADPH 氧化酶、p47phox 【Abstract】 Scopoletin （6-methoxy-7-hydroxycoumarin） is one of the most important phenolic compounds in noni. Scopoletin is a phenolic coumarin. Several studies have shown that scopoletin has antioxidant potential, antibacterial, anti-flammatory, and hepatoprotective activity effects. Scopoletin showed the relaxation of vascular to achieve hypotensive effect. Reactive oxygen species ROS play important roles in the pathogenesis of cardiovascular disease. Previous studies indicated that phorbol 12-myristate 13-acetate PMA stimulates ROS production in vascular smooth muscle cells VSMCs . PMA is a potent nonphysiological agonist used to activate NADPH oxidase. NADPH oxidase is a multi-subunit enzyme. NADPH oxidase comprises at least five components: P47phox、NoxA1 and Rac, p22phox, and Nox1. On stimulation, p47phox becomes phosphorylated and the cytosolic subunits form a complex that translocates to the membrane. The aim of this study was to investigate the effects of scopoletin on ROS generation in VSMCs. The result showed that fermented noni juice contains higher quantities on scopoletin. Structural resolution and mole