食品科学 ›› 2017, Vol. 38 ›› Issue (13): 174-181.doi: 10.7506/spkx1002-6630-201713029

• 营养卫生 • 上一篇    下一篇

Ang Ⅱ诱导人脐静脉内皮细胞建立高血压损伤模型

刘国艳,孙贝贝,张 洁,于苏宁,蒋栋磊,方维明,徐 鑫   

  1. 扬州大学食品科学与工程学院,江苏 扬州 225127
  • 出版日期:2017-07-15 发布日期:2017-07-11

Establishment of Angiotensin Ⅱ-Induced Model in Human Umbilical Vein Endothelial Cells

LIU Guoyan, SUN Beibei, ZHANG Jie, YU Suning, JIANG Donglei, FANG Weiming, XU Xin   

  1. College of Food Science and Engineering, Yangzhou University, Yangzhou 225127, China
  • Online:2017-07-15 Published:2017-07-11

摘要: 目的:采用高血压发病因子血管紧张素Ⅱ(angiotensin Ⅱ,Ang Ⅱ)诱导人脐静脉内皮细胞(humanumbilical vein endothelial cells,HUVECs)建立高血压损伤模型。方法:使用不同浓度(0.01、0.10、1.00 μmol/L和10.00 μmol/L)Ang Ⅱ诱导HUVECs不同时间(3、6、9、12 h和24 h),通过考察HUVECs形态、活性、功能、微观结构及凋亡程度来评价HUVECs损伤程度,同时采用交叉设计方差分析和多重比较方法得到Ang Ⅱ最适诱导浓度和最佳诱导时间。结果:Ang Ⅱ呈浓度依赖式诱导HUVECs损伤,最佳诱导条件为1.00 μmol/L Ang Ⅱ诱导12 h,此时HUVECs活性为44.85%、NO含量为43.57 μmol/L、总一氧化氮合酶活力为6.99 U/mg pro、内皮型一氧化氮合酶活力为1.89 U/mg pro、丙二醛含量为7.46 nmol/mL、超氧化物歧化酶活力为27.29 U/mg pro、细胞凋亡率为41.5%,微观结构显示核膜皱缩,核仁消失,胞浆内出现大量空泡状结构,线粒体或细小或肿胀,粗面内质网扩张数目较少,部分核糖体丢失,平面内质网扩张,但细胞未解体,仍然保持细胞结构。结论:1.00 μmol/L Ang Ⅱ诱导12 h可以成功诱导HUVECs建立高血压损伤模型。

关键词: 血管紧张素Ⅱ(Ang Ⅱ), 人脐静脉内皮细胞(HUVECs), 高血压, 细胞模型

Abstract: Objective: This research was aimed to establish a hypertensive damage model in human umbilical vein endothelial cells (HUVECs) induced by angiotensin (Ang) Ⅱ. Methods: HUVECs were induced with different concentrations (0.01, 0.10, 1.00 and 10.00 μmol/L) of Ang Ⅱ for different times (3, 6, 9, 12 and 24 h), respectively. The injury degree of HUVECs was evaluated by cellular morphology, viability, function, microstructure, and apoptosis. The optimal concentration and time were obtained by using variance analysis and multiple comparison tests. Results: Ang Ⅱ induced injury in HUVECs in a dose- and time-dependent manner. The optimal Ang Ⅱ concentration and induction time were respectively 1.00 μmol/L, and 12 h. After 12 h induction by 1 μmol/L Ang Ⅱ induced, the cell viability was 44.85%, nitric monoxide (NO) content was 43.57 μmol/L, total nitric oxide synthase (TNOS) activity was 6.99 U/mg pro, endothelial nitric oxide synthase (eNOS) activity was 1.89 U/mg pro, malonaldehyde (MDA) content was 7.46 nmol/mL, superoxide dismutase (SOD) activity was 27.29 U/mg pro, and apoptosis percentage was 41.5%. The microstructure showed that the nuclear membrane shrivel and nucleolus disappeared, and a large number of air bubbles appeared inside the cytoplasm. Small and swollen mitochondria, only a small number of expanded rough endoplasmic reticula, partial loss of ribosome, and endoplasmic reticulum expansion were observed, but the cells did not collapse, and still maintained their normal structure. Conclusion: Induction with 1.00 μmol/L Ang Ⅱ for 12 h allows successful establishment of a hypertensive damage model in HUVECs.

Key words: angiotensin Ⅱ (Ang Ⅱ), human umbilical vein endothelial cells (HUVECs), hypertension, cell model

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