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

作　　者：左的于[1] 夏勇[2] 石云敏 皮强中 罗素新[1] Zuo Deyu;Xia Yong;Shi Yunmin;Pi Qiangzhong;Luo Suxin(Division of Cardiology, The First Affliated Hospital of Chongqing Medical University;Davis Heart and Lung Research Institute , Division of Cardiovascular Medicine of Department of Molecular and Cellular Biochemistry, The Ohio State University College of Medicine, Columbus)

机构地区：[1]重庆医科大学附属第一医院心血管内科,重庆400016 [2]美国俄亥俄州立大学心肺研究所,美国哥伦布43210

出　　处：《重庆医科大学学报》2019年第3期255-259,共5页Journal of Chongqing Medical University

基　　金：国家重点基础研究发展计划(973计划)资助项目(编号:2014CB542402);国家自然科学基金资助项目(编号:81270210);重庆市科委资助项目(编号:cstc2015shmszx120066)

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、5μmol/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5μmol/L,检测其内皮型一氧化氮合酶(endothelial nitric oxide synthase,e NOS)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5μmol/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、5μmol/L的14,15-DHET处理BAECs后细胞内e NOS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5μmol/L的14,15-DHET处理BAECs 4、8、12、24 h后细胞内e NOS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5μmol/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e NOS蛋白表达,并且有促进血管形成的作用。Objective:To investigate the effect of 14,15-DHET on the expression of endothelial nitric oxide synthase(e NOS)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 e NOS was measured. Human umbilical vein endothelial cells(HUVECs)were divided into control group and treatment group,with a concentration 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 e NOS 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 e NOS protein in BAECs of 1.521 ±0.021,1.922±0.061,3.642±0.173,and 4.813±0.134,respectively,which was significantly higher than that in the control group(P <0.05). After HUVECs were stimulated by 5 μmol/L14,15-DHET,the treatment group had significantly longer total tubular length and total tubular branching length than the control 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 and0.035). Conclusion:The results of this study show that 14,15-DHET can increase the expression of e NOS in BAECs and promote angiogenesis.

关 键 词：14 15-二羟基二十碳三烯酸 牛主动脉内皮细胞 人脐静脉内皮细胞 

分 类 号：R331.3[医药卫生—人体生理学]