摘要目的:明确高尿酸血症是否通过诱导内皮型一氧化氮合酶（eNOS）脱偶联导致内皮细胞发生氧化应激造成内皮损伤。方法:（1）体外培养人脐静脉内皮细胞（HUVEC），以不同浓度的尿酸（0、200、400、600、800 μmol/L）作用24 h、48 h和72 h，DHE荧光染色法测定ROS水平，NO试剂盒测定上清液NO含量，观察尿酸对内皮细胞氧化应激的影响。（2）选择结果较稳定的600 μmol/L尿酸浓度，加或不加eNOS抑制剂L-NAME，作用48 h，上述方法测定ROS水平及NO含量，荧光定量PCR检测eNOS mRNA表达，观察高尿酸对eNOS脱偶联的影响。结果:高浓度的尿酸可以增加细胞内ROS水平，降低NO含量及eNOSmRNA表达。加入eNOS抑制剂L-NAME后，显著降低由高尿酸所引起的细胞内ROS水平并增加NO含量及eNOSmRNA表达。结论:高尿酸血症可以通过eNOS脱偶联使内皮细胞发生氧化应激从而导致内皮损伤。

abstractsObjective:To explore whether hyperuricemia causes endothelial dysfunction through oxidative stress induced by eNOS uncoupling.Methods:(1) To observe the influence of uric acid on endothelial cell oxidative stress, human umbilical vein endothelial cells(HUVECs) were stimulated with different concentrations of uric acid (0, 200, 400, 600, and 800 μmol/L) for 24, 48, and 72 h, respectively. Endothelial ROS was measured by ROS-sensitive fluorescent dye dihydroethidium (DHE). The content of NO was measured by chemiluminescence using NO test kit. (2) To observe the effects of high uric acid on eNOS uncoupling, HUVECs were incubated with uric acid (600 μmol/L) with or without L-NAME (eNOS inhibitor) for 48 hours. Endothelial ROS and the content of NO were measured by the above-mentioned methods. The level of eNOSmRNA was measured by real-time PCR.Results:High uric acid significantly reduced cell activity, increased ROS production, and decreased the NO levels released from HUVECs and the level of eNOSmRNA. Treatment with eNOS inhibitor L-NAME significantly ameliorated ROS level induced by the high level of uric acid, and improved the content of NO and the level of eNOSmRNA.Conclusions:Hyperuricemia can induce endothelial dysfunction by activating oxidative stress induced by eNOS uncoupling.