异丙酚对缺氧损伤人血管内皮细胞蛋白激酶C 表达及钙离子浓度的影响

中国临床药理学与治疗学 ›› 2007, Vol. 12 ›› Issue (4): 444-447.

异丙酚对缺氧损伤人血管内皮细胞蛋白激酶C 表达及钙离子浓度的影响

祖剑宇¹, 刁玉刚², 刘洁³, 陈卫民¹

¹中国医科大学附属盛京医院麻醉科, 沈阳110003, 辽宁;
²解放军第201 医院麻醉科, 辽阳111000, 辽宁;
³中国医科大学实验技术中心, 沈阳110001, 辽宁

收稿日期:2006-12-14 修回日期:2006-12-14 发布日期:2020-10-30
通讯作者: 陈卫民,男, 教授, 博士生导师, 研究方向:围术期器官保护。
作者简介:祖剑宇, 女, 医学博士, 主治医师, 研究方向:麻醉生理与药理学研究。E-mail:xy.zu @163.com

Effects of propofol on expression of protein kinase C and calcium ion concentration in human vascular endothelial cells induced by hypoxia injury

ZU Jian-yu¹, DIAO Yu-gang², LIU Jie³, CHEN Wei-min¹

¹Department of Anesthesiology, Second Affiliated Hospital, China Medical University, Shenyang 110003, Liaoning,China;
²Department of Anesthesiology, ²01 Hospital of PLA, Liaoyang 111000, Liaoning, China;
³Experiment Technique Center, China Medical University, Shenyang 110001, Liaoning, China

Received:2006-12-14 Revised:2006-12-14 Published:2020-10-30

摘要/Abstract

摘要： 目的探讨异丙酚对缺氧损伤所致人血管内皮细胞凋亡、细胞内钙稳态和蛋白激酶C(PKC) 表达的影响。方法依文献方法进行人脐静脉内皮细胞原代和继代培养, 建立人脐静脉内皮细胞缺氧复氧损伤模型。将培养至融合状态的细胞随机分组:C组(正常对照组) 、HR 组(缺氧30 min 后复氧6 h组) 和PR 组(异丙酚25 、50 、100 μmol L 组) 。采用流式细胞仪检测细胞凋亡, Western-blot 测定PKC 蛋白的表达,Meta Flour 单细胞内钙测定系统测定细胞内钙浓度变化。结果 PR 组细胞凋亡降低, 与HR 组(14.7±1.0) %比较差异有统计学意义(P<0.01),PR 50 μmol L 组(9.4±0.6) %、PR 100 μmol L 组(9.5±0.6) %与PR 25 μmol L 组(12.3±0.7) %比较差异有统计学意义(P<0.01);PR 组细胞内钙离子浓度明显降低, 与HR 组(117.3±6.0) %比较差异有统计学意义(P<0.01), PR 50 μmol L 组(48.2±4.4) %、PR 100 μmol L 组(47.0±6.3) %与PR25 μmol L 组(80.3±7.6) %比较差异有统计学意义(P<0.01);PR 组PKC 蛋白表达升高, 与HR 组(6.3±0.8) %比较差异有统计学意义(P<0.01), PR50 μmol L 组(27.0±2.4) %、PR 100 μmol L 组(27.5±2.6) %与PR 25 μmol L 组(19.0±1.7) %比较差异有统计学意义(P<0.01) 。结论缺氧复氧导致血管内皮细胞凋亡增加, 与抑制PKC 表达、升高细胞内钙离子浓度有关。异丙酚抑制缺氧复氧所致的人血管内皮细胞凋亡, 可能与其活化细胞内PKC, 降低钙超载, 维持细胞内钙稳态有关。

关键词: 异丙酚, 内皮细胞, 缺氧, 细胞凋亡

Abstract: AIM: To investigate the effects of propofol on the expression of protien kinase C and intracellular calcium homeostasis in human vascular endothelial cells induced by hypoxia-reoxygenation injury. METHODS: The endothelial cells were separated from human umbilical veins.The cells were cultured in normal medium or the medium containing different concentrations of propofol. The cells were further incubated in hypoxic condition for 30 min and then in normal condition for 6 h to induce hypoxia-reoxygenation injury.The cells were divided into groups:the control group (C group), the hypoxia-reoxygenation group (HR group) and propofol groups (PR 25, 50, 100 μmol L groups).The expression of PKC and the concentration of intracellular calcium were measured by Western-blot analysis and immunofluorescence method, respectively. RESULTS: The number of apoptosis increased after reoxygenation (14.7±1.0 %).Propofol inhibited the apoptosis significantly (P<0.01), and the inhibited effect of propofol on 50 μmol L(9.4±0.6) % and 100 μmol L(9.5±0.6) %was more significant compared with the propofol on 25 μmol L(12.3±0.7) % (P<0.01).The expression of PKC was significantly inhibited after reoxygenation (P<0.01).The concentration of intracellular calcium induced by hypoxia-reoxygenation injury was up-regulated (117.3±6.0) %.Propofol attenuated the concentration of intracellular calcium significantly (P<0.01) and was more effective at 50 and 100 μmol L (P<0.01).Propofol at a concentration of 25 μmol L could significantly (19.0±1.7) %upregulate the expression of PKC (P<0.01).Propofol at concentrations of 50 μmol L (27.0±2.4) % and 100 μmol L (27.5± 2.6) % showed a similar result but is significant compared with PR25 group (P<0.01). CONCLUSION: Hypoxia-reoxygenation injury induces apoptosis, which may be related to inhibiting the expression of PKC and increasing the concentration of intracellular calcium.Propofol inhibits the apoptosis of endothelial cells induced by hypoxia-reoxygenation injury, and the mechanism is possibly related to the activation of PKC and depression of calcium overload.

Key words: propofol, endothelial cells, anoxia, apoptosis

中图分类号:

R614
R965.2

祖剑宇, 刁玉刚, 刘洁, 陈卫民. 异丙酚对缺氧损伤人血管内皮细胞蛋白激酶C 表达及钙离子浓度的影响[J]. 中国临床药理学与治疗学, 2007, 12(4): 444-447.

ZU Jian-yu, DIAO Yu-gang, LIU Jie, CHEN Wei-min. Effects of propofol on expression of protein kinase C and calcium ion concentration in human vascular endothelial cells induced by hypoxia injury[J]. Chinese Journal of Clinical Pharmacology and Therapeutics, 2007, 12(4): 444-447.

参考文献

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