Korean J Lab Med.  2008 Oct;28(5):325-331. 10.3343/kjlm.2008.28.5.325.

Assessment of Hemorheological Deformability of Human Red Cells Exposed to tert-Butyl Hydroperoxide, Verapamil and Ascorbate by Ektacytometer

• Kim DH
• Kim YK
• Won DI
• Shin S
• Suh JS

• KMID: 1781563
• DOI: http://doi.org/10.3343/kjlm.2008.28.5.325

Abstract

BACKGROUND: Normal erythrocyte is deformable and this facilitates blood flow in the capillaries. Oxidative stress reduces the deformability of erythrocytes, and influences on blood flow in microcirculation. The objective of this study was to investigate the deformability of erythrocytes exposed to oxidative stress, the protective effects of verapamil and ascorbic acid against oxidative damages in erythrocytes, and the value of the microfluidic ektacytometer, RheoScan-D (RheoMeditech, Korea) in clinical application.
METHODS
Effects of oxidative stress on erythrocytes were investigated using tert-butyl hydroperoxide (tBHP). Before exposure to tBHP, the erythrocytes were pretreated with verapamil and ascorbic acid to examine their protective effect against oxidative damages. The deformability of erythrocytes was measured by the microfluidic ektacytometer, RheoScan-D.
RESULTS
When treated with tBHP, the deformability of erythrocytes was decreased (P<0.01) and methemoglobin (metHb) formation and mean corpuscular volume (MCV) of erythrocytes were increased (P<0.01, P<0.05) compared to those of the untreated control cells. Compared to the tBHP treated cells, pretreatment with verapamil increased the deformability of erythrocytes (P<0.01) and decreased metHb formation (P<0.01) and MCV (P<0.05). Likewise, pretreatment with ascorbic acid increased the deformability of erythrocytes (P<0.01) and decreased metHb formation (P<0.01).
CONCLUSIONS
Oxidative stress reduces the deformability of erythrocytes and the deformability could be one of markers for oxidative damage. Verapamil and ascorbic acid have protective role against tBHP induced oxidative stress. The ektacytometer, RheoScan-D used in this study is convenient for clinical measurement and could be used in various fields of clinical medicine.

Keyword

tert-butyl hydroperoxide; Erythrocyte deformability; Ektacytometer

MeSH Terms

Adult
Antioxidants/pharmacology
Ascorbic Acid/*pharmacology
Calcium Channel Blockers/pharmacology
Erythrocyte Deformability/*drug effects
Erythrocytes/drug effects/physiology
Humans
Male
Microfluidic Analytical Techniques/*instrumentation
Oxidative Stress
Statistics, Nonparametric
Verapamil/*pharmacology
tert-Butylhydroperoxide/*pharmacology

Figure

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  Fig. 1. Schematic diagram of RheoScan-D ektacytometer.

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  Fig. 2. Changes in red blood cell images as differential pressure.

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  Fig. 3. Changes in red blood cell images as 20 Pa (left), 0.3 Pa (right) and EI (=X-Y/X+Y).

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