非诺贝特促进人内皮细胞内皮一氧化氮合酶复偶联的作用研究

目的探讨非诺贝特能否对脂多糖（LPS）诱导的血管内皮一氧化氮合酶（eNOS）脱偶联发挥保护作用。方法体外培养人脐静脉内皮细胞（HUVECs）,用非诺贝特预处理HUVECs 2 h,再与LPS共孵育24 h,采用高效液相色谱法检测细胞四氢生物蝶呤（BH4）的表达水平,ELISA检测细胞eNOS表达水平和细胞上清一氧化氮（NO）浓度,利用Confocal方法检测细胞内活性氧（ROS）产生水平。结果与对照组比较,单纯LPS刺激组内皮细胞BH4表达水平降低,伴有eNOS表达下调和NO水平降低,而内皮细胞内ROS产生增加（P均〈0.05）。与单纯LPS刺激组比较,非诺贝特预处理组内皮细胞BH4表达水平升高,同时伴有eNOS表达上调和NO水平增加,而内皮细胞内ROS产生降低（P均〈0.05）。结论非诺贝特通过上调BH4水平,对LPS诱导的血管内皮细胞eNOS脱偶联有逆转作用,这可能是其发挥血管内皮保护作用的机制之一。

Fenofibrate re-couples endothelial nitric oxide synthase by restoring the level of tetrahydrobiopterin in human umbilical vein endothelial cells

Objective To investigate whether fenofibrate could re-couple endothelial nitric oxide synthase（eNOS）.Methods Human umbilical vein endothelial cells（HUVECs） were treated with fenofibrate for 2 hours,after that the cells were incubated with lipopolysaccharide（LPS） for 24 hours.The amount of intracellular tetrahydrobiopterin（BH4） was measured by high-performance liquid chromatography.The levels of intracellular eNOS and cell culture supernatant nitric oxide（NO） were measured by ELISA kits.A confocal laser scanning microscope was used to detect the level of intracellular reactive oxygen species（ROS）.Results Compared with control group,the levels of BH4,eNOS and NO were lower in LPS group,and ROS level was higher（P0.05）.The levels of BH4,eNOS and NO in pretreated group with fenofibrate were significantly higher than those in LPS group（P0.05）,the level of ROS was lower than that in LPS group（P0.05）.Conclusion Fenofibrate may protect against atherosclerosis by promoting the re-coupling of eNOS with increasing the level of BH4.