Determination of erythrocyte deformability and its correlation to cellular ATP release using microbore tubing with diameters that approximate resistance vessels in vivo
Determination of erythrocyte deformability and its correlation to cellular ATP release using microbore tubing with diameters that approximate resistance vessels in vivo
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DOI：
10.1039/b308225n
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发表时间：
2003-01-01
期刊：
影响因子：
4.2
通讯作者：
Spence, DM
中科院分区：
文献类型：
--
作者：
Fischer, DJ;Torrence, NJ;Spence, DM
描述了一种用于测量红细胞（RBC）在其直径近似于体内所遇到的力的管道中的变形性的新方法。在此，将来自兔的RBC加载到75 mm内径的50 cm部分的微孔管中，并连接到注射泵。然后将这段管连接到一段15 cm的25 μ m id管上。当缓冲液被泵送通过流动系统时，RBC从管道的两个部分被排空。然而，红细胞不能自由移动通过25 μ m的管段，导致细胞在该管段的入口处积聚。由注射泵输出的持续力导致RBC变形，直到所有细胞最终从流动系统中排出。发现测量达到最大压力的一半（1/2 P（max））所需的时间可用作RBC变形性的指标。对于给定的样品，简单的缓冲液导致达到1/2 P（最大值）的时间（6.9 +/- 0.2 s）比可变形RBC（21.6 +/- 0.8 s）更少。为了验证达到1/2 P（最大值）的时间增加确实是由于RBC，研究了RBC样本的各种血细胞比容，如预期的那样，发现1/2% RBC血细胞比容与7%血细胞比容（19.1 +/- 0.3 s）相比具有更高的1/2 P（最大值）值（26.0 s +/- 2.2 s）。此外，经戊二醛化学硬化的RBC比正常RBC的变形性低25%。最后，进行了一项研究，以检查RBC变形性和ATP释放之间的关系，发现ATP释放增加作为RBC变形性的函数。这种方法大大简化了变形性的测量，只采用注射泵和微孔管，并可能导致更完整的理解红细胞变形性的生理意义。
A novel method is described for measuring the deformability of red blood cells (RBCs) in tubing whose diameters approximate forces encountered in vivo. Here, RBCs from rabbits are loaded into a 50 cm section of 75 mm id microbore tubing and connected to a syringe pump. This section of tubing is then connected to a 15 cm section of 25 mum id tubing. As buffer is pumped through the flow system, the RBCs are evacuated from both sections of tubing. However, the inability of the RBCs to move freely through the 25 mum id section of tubing results in a buildup of cells at the inlet of this portion of tubing. The continued force output by the syringe pump results in a deformation of the RBCs until all of the cells are eventually evacuated from the flow system. It was found that a measurement of the time required to reach half of the maximum pressure (1/2P(max)) may be used as an indicator of the RBC deformability. For a given sample, a simple buffer results in less time to reach 1/2P(max) (6.9 +/- 0.2 s) than deformable RBCs (21.6 +/- 0.8 s). To verify that the increased amount of time to reach 1/2P(max) is indeed due to the RBCs, various hematocrits of an RBC sample were investigated and, as expected, it was found that a 12% RBC hematocrit had a higher 1/2P(max) value (26.0 s +/- 2.2 s) when compared to a 7% hematocrit (19.1 +/- 0.3 s). In addition, RBCs chemically stiffened with glutaraldehyde were shown to be 25% less deformable than normal RBCs. Finally, a study was performed to examine the relationship between RBC deformability and ATP release and it was found that ATP release increased as a function of RBC deformability. This method greatly simplifies deformability measurements, employing only a syringe pump and microbore tubing, and may lead to a more complete understanding of the physiological significance of erythrocyte deformability.