灯盏花素对人脐静脉内皮细胞胞内Ca^(2+)水平的调控作用

Effects of Breviscapine on intracellular Ca^(2+) concentration in human umbilical vein endothelial cells

目的探讨灯盏花素对培养的人脐静脉内皮细胞(HUVECs)胞内Ca2+水平([Ca2+]i)的调控作用。方法采用新一代Ca2+荧光探针Fluo-3/AM标记培养的HUVECs,激光共聚焦显微镜检测细胞胞内钙荧光信号,观察灯盏花素对培养的HUVECs胞内Ca2+水平的调控作用。结果在胞外有Ca2+或无Ca2+的情况下,灯盏花素均可引起[Ca2+]i的短暂性升高;灯盏花素的Ca2+释放作用与钙泵抑制剂CPA存在着交迭;灯盏花素能够抑制由KCl所引起的[Ca2+]i的升高;灯盏花素对胞内Ca2+池耗竭后胞外复Ca2+所引起的钙内流无明显阻断作用。结论灯盏花素可引起胞内Ca2+池的Ca2+释放,其释放的Ca2+来自CPA敏感的Ca2+池。灯盏花素也可抑制经电压依赖性Ca2+通道的Ca2+内流,对Ca2+池耗竭后引起的Ca2+内流通道无明显阻断作用。

Aim To investigate the effects of Breviscapine（BE） on intracellular Ca^2＋ concentration in human umbilical vein endothelial cells （HUVECs）. Methods HUVECs were incubated with the calcium ion-sensitive fluorescent indicator fluo-3/AM, and then a laser confocal microscope was applied to measure changes of fluorescence intensity under different agonists to investigate the effects of Breviscapine on intracellular Ca^2＋ concentration in HUVECs. Results （1） BE induced Ca^2＋ transients in the buffer with or without Ca^2＋ ;（2）The Ca^2＋ transients induced by BE could be overlapped by cyclopiazonic acid （CPA） ;（3）Pretreatment of BE inhibited Ca^2＋ influx caused by KC1. （4） BE had no evident effect on the calcium influx indueed by the restoration of extraeellular Ca^2＋ after the depletion of intracellular Ca^2＋ stores. Conclusion BE depletes CPA-sensitive intracellular Ca^2＋ stores and inhibits Ca^2＋ influx through voltage-dependent Ca^2＋channels, and BE influx induced by has no evident effect on the calcium the restoration of extracellular Ca^2＋ after the depletion of intracellular Ca^2＋ stores.