高等学校化学学报

高等学校化学学报 ›› 2002, Vol. 23 ›› Issue (6): 985.

• 研究论文 •    下一篇

稀土离子跨人血红细胞膜的荧光法研究

杨频, 魏春英, 陈娟   

  1. 山西大学分子科学研究所, 太原 030006
  • 收稿日期:2001-08-02 出版日期:2002-06-24 发布日期:2002-06-24
  • 通讯作者: 杨　频(1933年出生),男,教授,博士生导师,从事生物无机化学及配位化学研究.E-mail:yangpin@sxu.edu.cn E-mail:yangpin@sxu.edu.cn
  • 基金资助:

    国家自然科学基金重大项目(批准号:29890280)资助

Studies on Trans-human Red Cells of Rare Earth Ions by Fura-2 Indicator

YANG Pin, WEI Chun-Ying, CHEN Juan   

  1. Institute of Molecular Science, Shanxi University, Taiyuan 030006, China
  • Received:2001-08-02 Online:2002-06-24 Published:2002-06-24

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被引次数

37

摘要： 采用Fura-2荧光浓度指示剂对红细胞的稀土跨膜作用进行了系列研究.结果表明,稀土离子不能通过完整的红细胞膜进入细胞内.通过与离子载体实验相对照,发现细胞ATP耗竭后,低浓度的稀土离子(5×10-6mol/L)不能跨膜进入ATP-耗竭红细胞.KCl去极化及加入电压依赖性钙通道刺激剂Bay-K8644对稀土离子的跨膜也没有促进作用.在Ca2+内流正常的情况下,低浓度稀土离子(5×10-6mol/L)对钙离子内流无影响.增大稀土离子浓度到5×10-4mol/L,用显微镜观察此时红细胞已开始溶血.在模拟胞内离子组分的缓冲液中(pH=7.05),比较了La3+,Eu3+和Ca2+对Fura-2的敏感程度.此条件下Fura-2对La3+和Eu3+的检测限分别为10-12和10-14mol/L,对Ca2+的检测限为10-8mol/L,并测得Fura-2-La3+(Eu3+)的络合比为1∶1,表观离解常数为1.7×10-12和4.95×10-14mol/L,表明用此法检测稀土离子跨膜行为相当灵敏有效.

关键词: ATP-耗竭, 人血红细胞, Fura-2, 稀土离子

Abstract: The transmembrane behavior of La3+ and Eu3+ was examined by using Fura-2 technique. The result showing a low concentration La3+ or Eu3+ (5×10-6mol/L) could not enter fresh and ATPdepleted human red cells. But in the presence of ionomycin, obvious transmembrane phenomenon was observed when La3+ (or Eu3+)of 5×10-6mol/Lwas added. KCl and Bay K8644 could not promote the influx of La3+ (or Eu3+). Moreover, a low concentration La3+ (or Eu3+) had no effect on the influx of Ca 2+ in ATPdepleted human red cells. When a higher concentration La3+ or Eu3+ (5×10-4mol/L) was added, red cells became hemolysis. Acharacterization of La3+ (or Eu3+)-Fura-2 interactions showed that they formed a 1∶1 La3+ (or Eu3+)-Fura-2 complex. In vitro studies confirmed that Fura-2 fluorescence was sensitive to La3+ (or Eu3+) and the apparent dissociation constants of La3+ and Eu3+ for Fura-2 (Kd) were 1.7×10-12 and 4.95×10-14mol/L, respectively (pH=7.05).

Key words: ATP depleted, Human red cells, Fura-2, Rare earth ions

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