14，15-DHET对牛主动脉内皮细胞中内皮型 一氧化氮合酶表达影响的研究

Effect of 14，15-DHET on the expression of endothelial nitric oxide synthase in bovine aortic endothelial cells

摘要:

目的：探讨14，15-二羟基二十碳三烯酸（14，15-DHET）对牛主动脉内皮细胞中内皮型一氧化氮合酶表达以及血管形成的影响。方法：将牛主动脉内皮细胞（bovine aortic endothelial cells，BAECs）按照不同14，15-DHET处理浓度分组为0（对照组）、0.1、0.5、1、5 μmol/L组，刺激时间均为24 h；按照时间梯度分组为0（对照组）、4、8、12、24 h，刺激浓度均为5 μmol/L，检测其内皮型一氧化氮合酶（endothelial nitric oxide synthase，eNOS）的蛋白表达水平，同时将人脐静脉内皮细胞（human umbilical vein endothelial cells，HUVECs）按照不同14，15-DHET浓度分组为0（对照组）和5 μmol/L（处理组），刺激24 h后观察14，15-DHET对血管形成的影响。结果：在浓度梯度组中用0.1、0.5、1、5 μmol/L的14，15-DHET处理BAECs后细胞内eNOS蛋白表达水平（1.456±0.008、2.202±0.045、3.931±0.170、4.588±0.179）与对照组相比均明显升高（P＜0.05）；在时间梯度组中用5 μmol/L的14，15-DHET处理BAECs 4、8、12、24 h后细胞内eNOS蛋白表达水平（1.521±0.021、1.922±0.061、3.642±0.173、4.813±0.134）与对照组相比均明显升高（P＜0.05）；使用5 μmol/L的14，15-DHET刺激HUVECs后，处理组小管总长度和小管分枝总长度（13 715.000±448.092，12 538.000±574.182）与对照组（10 426.000±950.537，8 672.000±1 025.486）相比均明显升高（P=0.043，P=0.035）。结论：本研究结果表明14，15-DHET可以增加牛主动脉内皮细胞的eNOS蛋白表达，并且有促进血管形成的作用。

Abstract:

Objective：To investigate the effect of 14，15-DHET on the expression of endothelial nitric oxide synthase（eNOS） in bovine aortic endothelial cells（BAECs） and angiogenesis. Methods：According to the concentration of 14，15-DHET，BAECs were divided into 0（control group），0.1，0.5，1，and 5 μmol/L groups，with a stimulation time of 24 hours；according to the time gradient，BAECs were divided into 0（control group），4，8，12，and 24 hours groups，with a stimulation concentration of 5 μmol/L. The expression of eNOS was measured. Human umbilical vein endothelial cells（HUVECs） were divided into control group and treatment group，with a con－centration of 14，15-DHET of 0 and 5 μmol/L，respectively，and the effect of 14，15-DHET on angiogenesis was observed after 24 hours of stimulation. Results：As for concentration gradient，the 0.1，0.5，1，and 5 μmol/L 14，15-DHET groups had an expression level of eNOS protein in BAECs of 1.456±0.008，2.202±0.045，3.931±0.170，and 4.588±0.179，respectively，which was significantly higher than that in the control group（P<0.05）. As for time gradient，the 4，8，12，and 24 hours groups treated with 5 μmol/L 14，15-DHET had an expression level of eNOS protein in BAECs of 1.521±0.021，1.922±0.061，3.642±0.173，and 4.813±0.134，respec－tively，which was significantly higher than that in the control group（P <0.05）. After HUVECs were stimulated by 5 μmol/L 14，15-DHET，the treatment group had significantly longer total tubular length and total tubular branching length than the con－trol group（13 715.000±448.092 and 12 538.000±574.182 vs. 10 426.000±950.537 and 8 672.000±1 025.486，P=0.043 and 0.035）. Conclusion：The results of this study show that 14，15-DHET can increase the expression of eNOS in BAECs and promote angiogenesis.