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应用压电石英晶体生物传感器研究赤芍801与ET-1间的相互作用

Study on Interaction of Radix Paeoniae 801 and endothelin-1 by Using a Piezoelectric Quartz Crystal Biosensor

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【作者】 李静吴宝艳蒋跃绒殷惠军黄加栋陈可冀陈强

【Author】 LI Jing, WU Bao yan, JIANG Yue rong, et al Life Science College, Nankai University, Tianjin (300071)

【机构】 南开大学生命科学院中国中医研究院西苑医院南开大学生命科学院 天津300071天津300071天津300071

【摘要】 目的采用体外实验的方法 ,直接检测活血化瘀中药赤芍有效成分赤芍 80 1与细胞因子内皮素 1(ET 1)间的相互作用 ,探讨其作用的可能的靶点和分子机理。方法应用压电石英晶体生物传感器———石英晶体微天平 (QCM )检测赤芍 80 1与ET 1之间的特异性结合。在QCM金基片表面经活化后接合上亲和素 ,继而将生物素衍生化的ET 1固定于基片表面 ,加入赤芍 80 1,检测赤芍 80 1与ET 1之间的相互作用。每一步结合反应后 ,均用磷酸盐缓冲液 (PBS)冲洗以去除非特异性吸附。结果赤芍 80 1与ET 1发生特异性结合。结论ET 1可能是赤芍 80 1体内作用靶点之一 ,即赤芍 80 1通过与ET 1结合 ,阻碍ET 1与其受体间的结合 ,从而拮抗ET 1的作用 ,扩张血管 ,抑制血小板聚集。

【Abstract】 Objective To explore the possible target and molecular mechanism of Radix Paeoniae 801 (RP801), an effective ingredient extracted from Radix Paeoniae, the Chinese herbal medicine for activating blood circulation to remove blood stasis, using experimental in vitro method by directly detecting the interaction between RP801 and endothelin 1 (ET 1). Methods Piezoelectric quartz crystal biosensor, namely, the quartz crystal microbalance (QCM) was used to detect the specific combining between RP801 and ET 1 by binding avidin to the pre activated Au surface of electrode of QCM, followed by immobilizing biotinylated ET 1 to it, and adding RP801, then the binding curve was recorded. PBS washing was applied at the end of every steps of combining reaction for dissociate the non specific absorption. Results Specific combining of RP801 and ET 1 was found. Conclusion ET 1 could possibly be one of the acting targets of RP801 in the body, that is, RP801 could combine with ET 1 to impede the binding of ET 1 with its receptor, so as to counteract the action of ET 1, dilate blood vessels and inhibit platelet aggregation.

【基金】 国家自然科学基金资助项目 (No .90 2 0 90 54)
  • 【文献出处】 中国中西医结合杂志 ,Chinese Journal of Integrated Traditional and Western Medicine , 编辑部邮箱 ,2004年08期
  • 【分类号】R285
  • 【被引频次】21
  • 【下载频次】163
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