摘要
目的 研究主动脉内皮细胞乙酰胆碱激活蛋白(EPA)介导胞内钙离子变化的特征 ,并探讨培养牛主动脉内皮细胞EPA功能的变化。方法 以Fluo 3,Frua 2为荧光探针 ,采用共聚焦和荧光光度法测定内皮细胞 [Ca2 + ]i。结果 Ach能够引起体外培养 6代以内内皮细胞 [Ca2 + ]i 的增加 ;Ach激发的内皮细胞 [Ca2 + ]i 信号表现为瞬时性单次或连续 2次的钙振荡 ,具有异时性和快速耐受的特点。未观察到烟碱和其它M受体激动剂引起的 [Ca2 + ]i 变化。Ach对胞内 [Ca2 + ]i 浓度的作用以原代细胞增加最为明显 ,但随培养时间的延长 ,此效应逐渐减弱 ,至 13代后无效应。结论 血管内皮细胞乙酰胆碱激活蛋白激活能介导胞内钙离子的增高 ,经培养后EPA功能丧失。
AIM To examine the change of intracelluar Ca 2+ level regulated by EPA in bovine aorta endothelium of different stages of culture and determine the time that cultured endothelium loses functional EPA. METHODS As fluorescence probe, Fluo 3 and Fura 2 were used in assaying [Ca 2+ ] i in endothelium by confocal microscopy and fluorophotometry. RESULTS Ach could activate EPA and elicite the [Ca 2+ ] i change of endothelium in certain periods of culture, especially in primary stage of cul ture. The rises of [Ca 2+ ] i exhibited as oscillation characterized by iso timing and immediate tolerance. The functional EPA was lost in passage 13. CONCLUSION EPA can be activated by Ach, which mediated the rise of intracelluar Ca 2+ in bovine aorta endothelia cell. This character of EPA is lost during passaging.
出处
《中国药理学通报》
CAS
CSCD
北大核心
2001年第3期275-278,共4页
Chinese Pharmacological Bulletin
基金
国家重点基础研究发展规划项目!资助课题
NoG .19980 5 1112
