摘要
目的 通过观察猪主动脉内皮细胞在血管紧张素Ⅱ（AngⅡ）作用下时，不同浓度和不同作用时间的丹参酮ⅡA（TSN）对血管内皮细胞分泌一氧化氮（NO）及其内皮型一氧化氮合酶（eNOS）基因表达的影响、细胞内游离钙离子浓度的变化，探讨TSN对血管内皮细胞的保护作用。方法采用硝酸还原法、免疫组织化学法，首先分别检测不同浓度（10^-8～10^-6mol／L）和不同作用时间（1、6、24h）的AngⅡ对培养的猪主动脉内皮细胞产生NO及其eNOS蛋白质表达的影响；然后比较在10^-6mol／L的AngII作用的不同点（0h点为A组、6h点为B组）加入不同浓度（10、50mg／L）TSN，分别检测作用1、6、24h后内皮细胞的NO生成和eNOS的蛋白质表达变化。用激光共聚焦扫描显像系统检测内皮细胞内游离钙离子浓度（[-Ca^2＋]i）水平的变化。结果 （1）随着AngⅡ浓度的增加、作用时间的延长，内皮细胞NO的产生及eNOS的表达呈顺序下降（P〈O．01），表现出剂量、时间依赖性的负性作用。（2）TSN可抑制AngⅡ对内皮细胞分泌NO及eNOS表达的负性作用（P〈0．01），但尚不能恢复到空白对照组水平（P〈0．05）；此作用与TSN的浓度无明显关系（P〉0．05）。在TSN作用1、6h，A组的抑制效应明显强于B组（P〈0．05）；随着作用时间延长至24h，两组间差异无显著性（P〉0．05）。（3）AngⅡ可引起主动脉内皮细胞[-Ca^2＋]i显著升高（P〈0．01），TSN可部分抑制AngII诱导的内皮细胞[-Ca^2＋]i升高（P〈0．05）。结论 TSN可抑制AngⅡ对血管内皮细胞分泌NO以及细胞eNOS蛋白质表达的负性作用，可能通过多种途径对血管内皮细胞及其功能起到保护作用。
Objective To investigate the protective effect of tanshinone Ⅱ A on changes in nitric oxide （NO） level and endothelial nitric oxide synthase [eNOS） gene expression induced by angiotensin Ⅱ [Ang Ⅱ ） in porcine aortic endothelial cells [PAEC）. Methods The effect of different concentration [10^-8-10^-6mol/L） and effect time [1 h, 6 h and 24 h） of Ang Ⅱ on NO level and expression of eNOS was measured by nitric acid deoxidizing and immunohistochemistry methods. We cocultured endothelial cells with tanshinone Ⅱ A and studied its role on NO level and eNOS protein expression at 1 h, 6 h and 24 h after Ang Ⅱ 10^-8 mol/L was added, [-Ca^2＋]i was observed with laser-scanning confocal microscope. Results （1） With the increasing of concentration and effect time of Ang Ⅱ , the level of NO and eNOS in PAEC was decreased in dose and time-dependent manner [P〈 0.01）. （2）Tanshinone Ⅱ Ainhibited the decreased effect by Ang Ⅱ on NO and eNOS （P〈0.05）. The inhibition effect of tanshinone Ⅱ A on Ang Ⅱ was diminished with time. （3）Ang Ⅱ increase [Ca^2＋ ]i in PAEC, which was inhibited by tanshinone Ⅱ A. Conehtsion Tanshinone Ⅱ A have a protective role on vascular endothelial cells and its function by suppressing the inhibition of Ang Ⅱ effect on the level of NO and eNOS gene expression in PAEC.
出处
《中华高血压杂志》
CAS
CSCD
北大核心
2006年第11期882-886,共5页
Chinese Journal of Hypertension
基金
国家自然科学基金资助项目
项目批准号:30500657
