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晚期糖基化终末产物对人脐静脉内皮细胞分泌NO功能的影响

张胜雷,徐金升,王悦芬,白亚玲,崔立文,张慧然   

  1. 河北医科大学第四医院
  • 收稿日期:2012-08-30 修回日期:2013-01-18 出版日期:2013-06-15 发布日期:2013-06-15
  • 通讯作者: 徐金升

Effects of advanced glycation end products (AGEs) on the NO in the HUVECs

  • Received:2012-08-30 Revised:2013-01-18 Published:2013-06-15 Online:2013-06-15

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摘要： 【摘要】 目的  探讨晚期糖基化终产物（AGEs）对人脐静脉内皮细胞一氧化氮分泌功能的影响。方法  培养人脐静脉内皮细胞，制备晚期糖基化终末产物（AGEs-HSA）。将体外培养的人脐静脉内皮细胞随机分为AGEs-HSA组和HSA（人血白蛋白）组， 分别用不同浓度的AGEs-HSA、HSA刺激内皮细胞24 h。然后用酶联免疫法分别测定细胞培养上清液中的NO含量、NOS活性、SOD活性和MDA含量。结果  1与HSA组相比，AGEs-HSA组在浓度为100、200、400 mg/L 时能够明显抑制内皮细胞NO的产生和NOS的活性，并呈浓度依赖性（P <0.05）。2与HSA组相比，AGEs-HSA组在浓度为25、50、100、200、400 mg/L 时均能够明显抑制内皮细胞SOD的活性和促进MDA含量的增加，并呈浓度依赖性（P <0.05）。3 AGEs-HSA组中在AGEs-HSA刺激浓度为400 mg/L时， NO浓度和NOS活性、SOD活性呈正相关（P <0.01），而与MDA含量呈负相关（P <0.01）。NOS的活性与SOD活性呈正相关（P <0.01），与MDA含量呈负相关（P <0.01）。结论 晚期糖基化终末产物能够抑制人脐静脉内皮细胞NO的分泌。其机制之一可能是AGEs介导内皮细胞发生氧化损伤致使NOS活性降低进而导致内皮细胞NO分泌下降。

关键词: 糖基化终产物, 高级, 内皮细胞, 脐静脉, 一氧化氮, 一氧化氮合酶, 超氧化物歧化酶, 丙二醛

Abstract: [Abstract]   Objective   To investigate effects of advanced glycation end products (AGEs) modified protein on production of nitric oxide (NO) in cultured human umbilical vein endothelial cells (HUVECs). Methods   HUVECs were co-incubated with AGE-HSA, HSA for 24h at different concentrations. Nitric oxide production and activity of nitric oxide synthase in supernatant were measured by NO and NOS assay kits. The activity of SOD and contents of MDA were determined by SOD and MDA assay kits. Results  1 The contents of NO and activity of NOS in AGEs group were significantly lower than those in HSA group and in a concentration-dependent manner (P <0.05). 2 The activity of SOD was significantly lower than that in HSA group and in concentration – dependent manner (P <0.05). The concentration of MDA was significantly higher than that in HSA group and in concentration–dependent manner (P <0.05). 3 The concentrations of NO in supernatant were positively associated with the activity of NOS and SOD, respectively (P <0.01). The concentrations of NO in supernatant were negatively associated with the contents of MDA (P <0.01). The activity of NOS was positively associated with the activity of SOD (P <0.01). The activity of NOS was negatively associated with the contents of MDA (P <0.01). Conclusion   The oxidative damage of AGEs induced by the HUVECs may play an important role in the decrease of NO in the HUVECs.

Key words: endothelial cells, umbilical veins, nitric oxide, nitric oxide synthase, superoxide dismutase, malondialdehyde, 丙二醛