《中国康复理论与实践》

《中国康复理论与实践》 ›› 2008, Vol. 14 ›› Issue (05): 440-441.

• 基础研究 • 上一篇    下一篇

临床相关浓度硫喷妥钠对大鼠脑胶质瘤细胞核因子-κB活性的影响

范婷1; 安建雄1; 杜培革2   

  1. 1.清华大学玉泉医院麻醉与疼痛医学科,北京市 100049;2.北华大学医学院,吉林吉林市 132011
  • 收稿日期:2008-01-21 出版日期:2008-05-01 发布日期:2008-05-01

Effect of Thiopental on Nuclear Factor Kappa B in Rat Glioma Cells

FAN Ting, AN Jian-xiong, DU Pei-ge   

  1. Department of Anaesthesiology, Tsinghua Univeristy Yuquan Hospital, Beijing 100049, China
  • Received:2008-01-21 Published:2008-05-01 Online:2008-05-01

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摘要： 目的探讨临床相关浓度的硫喷妥钠及其不同作用时间对大鼠脑胶质瘤细胞核因子(NF)-κB活性的影响。方法取大鼠脑胶质瘤C6细胞株,分别与10-3mol/L、10-4mol/L、10-5mol/L硫喷妥钠作用1h,在收集细胞前30min用10ng/ml脂多糖进行处理;取大鼠脑胶质瘤C6细胞株,分别与10-5mol/L硫喷妥钠作用0.5h、1h、2h、4h,在收集细胞前30min用脂多糖进行处理。提取细胞总RNA,逆转录得到c-DNA,之后行PCR反应;NF-κB表达结果进行凝胶电泳分析(EMSA)并用Bio-Rad密度仪进行半定量分析。结果经10-3mol/L、10-4mol/L、10-5mol/L硫喷妥钠处理的电泳带窄于对照,密度测量值低于对照(P<0.05),但各处理浓度之间密度测量值的无显著性差异。经10-5mol/L硫喷妥钠作用0.5h、1h、2h、4h的各电泳带宽度依次递减,亮度依次递减;密度测量数值也依次递减(P<0.05),且低于对照(P<0.05)。结论硫喷妥钠对脑胶质瘤细胞NF-κB活性具有明显影响。这种影响与硫喷妥钠浓度无关,随硫喷妥钠作用时间的延长而增强。

关键词: 核因子-κB(NF-κB), 胶质瘤细胞, 硫喷妥钠, 浓度, 作用时间

Abstract: Objective To investigate the effect of thiopental on lipopolysaccharide(LPS)-induced activation of nuclear factor kappa B(NF-κB) under different concentration or after different time in rat glioma cells in vitro.Methods The rat C 6 glioma cells were treated with,in the presence or Absence of various concentrations(10-3 mol/L,10-4 mol/L and 10-5 mol/L)of thiopental for 1 h.30 min before harvest,the cells were exposed to LPS(10 ng/ml).The other C 6 cells were treated with 10-5 mol/L thiopental with different time(0.5 h,1 h,2 h,4 h).30 min before harvest,the cells were exposed to LPS(10 ng/ml).Nuclear extracts from C 6 cells were harvested using standard procedures,and the LPS-induced NF-κB expression was determined with electrophoretic mobility shift assays(EMSA) and measured with densitometry(Bio-Rad).Results The width of the bands of NF-κB under thiopental with different concentrations were similar,but they were narrower than that of the control(treated only with LPS),the density was less than that of the control(P<0.05).The width of the bands of NF-κB under thiopental with different time decreased with the time(P<0.05),and they were much narrower and shallower than that of the control(P<0.05),and the density was significantly less than that of the control(P<0.05).Conclusion Thiopental suppressed the activation of transcription factor NF-κB,and the inhibition was concentration-independent but time-dependent.

Key words: nuclear factor kappa B(NF-κB), glioma cell, thiopental, concentration, time