上海交通大学学报（医学版）

›› 2011, Vol. 31 ›› Issue (12): 1676-.doi: 10.3969/j.issn.1674-8115.2011.12.004

• 论著（基础研究） • 上一篇    下一篇

蛋白激酶C-ε在丙泊酚诱导内皮源性一氧化氮合成酶激活中的作用

祁爱花1, 王 莉2, 崔德荣2, 周全红2, 江 伟2   

  1. 1.苏州大学研究生部, 苏州 215006； 2.上海交通大学附属第六人民医院麻醉科, 上海 200233
  • 出版日期:2011-12-28 发布日期:2012-01-04
  • 通讯作者: 王 莉, 电子信箱: liwang1118@hotmail.com。
  • 作者简介:祁爱花（1984—）, 女, 硕士生；电子信箱: chinaqi@126.com。
  • 基金资助:

    国家自然科学基金（30972842）；上海市自然科学基金（09ZR1424000）

Role of protein kinase C-&epsilon|in activation of endothelial nitric oxide synthase induced by propofol

QI Ai-hua1, WANG Li2, CUI De-rong2, ZHOU Quan-hong2, JIANG Wei2   

  1. 1.Department of Postgraduate, Soochow University, Suzhou 215006, China；2.Department of Anesthesiology, the Sixth People's Hospital, Shanghai Jiaotong University, Shanghai 200233, China
  • Online:2011-12-28 Published:2012-01-04
  • Supported by:

    National Natural Science Foundation of China, 30972842；Shanghai Natural Science Foundation, 09ZR1424000

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摘要：

目的 在人脐静脉内皮细胞（HUVECs）模型中观察丙泊酚对内皮源性一氧化氮合成酶（eNOS）的激活效应，探讨蛋白激酶C-ε（PKC-ε）对药物引起eNOS激活的调节作用。方法 以体外培养的人HUVECs 3～5代作为实验对象。用丙泊酚（0、1、5、10、50、100 μmol/L）分别处理细胞1和24 h，分析药物激活eNOS的量效关系；用丙泊酚5 μmol/L和50 μmol/L分别处理细胞0 min、5 min、1 h、6 h和24 h，分析药物激活eNOS的时效关系。细胞随机分为：①正常对照组；②PKC-ε假底物+丙泊酚5 μmol/L组；③PKC-ε假底物+丙泊酚50 μmol/L组；④单独PKC-ε假底物组；⑤单独丙泊酚50 μmol/L组；⑥10%长链脂肪乳组。各组分别处理细胞1和24 h。Western blotting分析PKC-ε、eNOS、P-Ser1177-eNOS蛋白的表达。结果 丙泊酚呈剂量、时间依赖性上调P-Ser1177-eNOS蛋白的表达。处理细胞24 h时，与单独丙泊酚50 μmol/L组相比，PKC-ε假底物+丙泊酚50 μmol/L组P-Ser1177-eNOS蛋白的表达显著降低（P＜0.05)。结论 丙泊酚呈剂量、时间依赖性诱导eNOS的激活，PKC-ε参与介导药物对eNOS的激活效应。

关键词: 丙泊酚, 内皮源性一氧化氮合成酶, 蛋白激酶C-ε, 人脐静脉内皮细胞

Abstract:

Objective To observe the effect of propofol on endothelial nitric oxide synthase (eNOS) activation, and explore the regulation role of protein kinase C-ε (PKC-ε) in drug-induced eNOS activation in human umbilical vein endothelial cells (HUVECs) model. Methods HUVECs of the third to fifth passages cultured in vitro were selected. Cells were treated with propofol (0 μmol/L, 1 μmol/L, 5 μmol/L, 10 μmol/L, 50 μmol/L and 100 μmol/L) for 1 h and 24 h respectively, and the concentration-effect relationship of propofol in activation of eNOS was analysed. Cells were exposed to propofol (5 μmol/L and 50 μmol/L) for 0 min, 5 min, 1 h, 6 h and 24 h, and time-effect relationship of propofol in activation of eNOS was explored. Then, cultured HUVECs were randomly divided into normal control group, PKC-ε pseudosubstrate+ propofol 5 μmol/L group, PKC-ε pseudosubstrate+ propofol 50 μmol/L group, single PKC-ε pseudosubstrate group, single propofol 50 μmol/L group and 10% long chain fat emulsion group. Cells in each group were treated for 1 h and 24 h respectively. The expression of PKC-ε, eNOS and P-Ser1177-eNOS protein was detected by Western blotting. Results Propofol increased the expression of P-Ser1177-eNOS protein in concentration- and time-dependent manners. When cells were treated for 24 h, the expression of P-Ser1177-eNOS protein in PKC-ε pseudosubstrate+ propofol 50 μmol/L group was significantly lower than that in single propofol 50 μmol/L group （P＜0.05). Conclusion Propofol induces the activation of eNOS in concentration- and time-dependent manners, and PKC-ε positively mediates the activation of eNOS induced by propofol.

Key words: propofol, endothelial nitric oxide synthase, protein kinase C-ε, human umbilical vein endothelial cell