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目的:观察氟伐他汀对人脐静脉内皮细胞(HUVECs)游离钙离子水平及内皮型一氧化氮合酶(eNOS)活性的影响及可能机制。方法:体外培养HUVECs,随机分为5组:空白对照组,氟伐他汀(10-8,10-7,10-6,10-5mol/L)组。采用硝酸还原酶法测定细胞上清液中NO含量,液体闪烁计数仪测定L-[3H]-精氨酸和L-[3H]-瓜氨酸的含量,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([Ca2+]i)水平的变化。结果:与空白对照组比较,10-8,10-7,10-6,10-5mol/L氟伐他汀孵育细胞12h后可显著升高HUVECs细胞内eNOS活性,促进NO释放,同时伴有[Ca2+]i升高,且呈浓度依赖性。另外,10-5mol/L氟伐他汀在0～12h时间段呈时间依赖性增高eNOS活性,作用12h使eNOS活性达到最高(P<0.01)。结论:氟伐他汀呈浓度依赖性升高HUVECseNOS活性和促进NO释放,该作用与其增加内皮细胞内[Ca2+]i有关。
Objective: To observe the effect of fluvastatin on the level of free calcium and the activity of endothelial nitric oxide synthase (eNOS) in human umbilical vein endothelial cells (HUVECs) and its possible mechanism. Methods: HUVECs were cultured in vitro and randomly divided into 5 groups: blank control group, fluvastatin (10-8,10-7,10-6,10-5mol / L) group. The nitric acid reductase method was used to determine the content of NO in the cell supernatant. The content of L- [3H] - arginine and L- [3H] - citrulline was measured by liquid scintillation counter. The confocal laser scanning microscope Changes in intracellular free calcium concentration ([Ca2 +] i) levels. Results: Compared with the blank control group, the cells incubated with fluvastatin 10-8, 10-7, 10-6, 10-5mol / L for 12h significantly increased the intracellular eNOS activity and NO release in HUVECs, Ca2 +] i increased in a concentration-dependent manner. In addition, 10-5mol / L fluvastatin increased eNOS activity in a time-dependent manner at 0-12h, and reached the highest eNOS activity at 12h (P <0.01). Conclusion: Fluvastatin can increase the activity of HUVECseNOS and promote the release of NO in a concentration-dependent manner, which is related to the increase of [Ca2 +] i in endothelial cells.