Abstract

Osmotic fragility (OF) in red blood cells (RBCs) is a useful tool for evaluating the actions of various chemicals on the cell membrane in vitro. The effects of monocarboxylic and dicarboxylic acids on OF were evaluated in cattle RBCs. Isolated cattle RBCs were immersed in various carboxylic acids at 0-100 mM in a buffer solution for 1 hr and the 50% hemolysis was then determined by soaking in 0.1-0.8% NaCl solution. Although n-caprylic acid at 100 mM induced hemolysis, the other monocarboxylic acids possessing straight hydrocarbons did not affect OF. The dicarboxylic acids possessing straight hydrocarbons, except for glutaric acid, decreased OF in a dose-dependent manner. Some monocarboxylic acids with branched hydrocarbons tended to decrease OF, but these changes were not statistically significant. Although cyclopentanecarboxylic and cyclohexanecarboxylic acids at 100 mM decreased OF, other monocarboxylic acids with cyclic hydrocarbons did not affect OF. Among the dicarboxylic acids with cyclic hydrocarbons tested, only 1,2-cyclohexanedicarboxylic acid and phthalic acid with a benzene ring significantly decreased OF. There is no clear correlation between the effect of monocarboxylic or dicarboxylic acids on OF, and their partition coefficients. Thus, the partition coefficient is not a suitable parameter for explaining the effect of both groups of carboxylic acids on OF in cattle RBCs. With regard to the effect of monocarboxylic acids on OF, although an increase in OF was demonstrated in rat RBCs, no effect or rather a decrease in OF was demonstrated in cattle RBCs, similar to the results observed for guinea pig and sheep RBCs. With regard to the effect of dicarboxylic acids, decreases in OF were already demonstrated in rat, guinea pig and sheep RBCs. We have proposed that dicarboxylic acids exhibit a common stabilizing effect on the RBC membrane in various animals, which we termed a “wedge-like effect”. We clarified that cattle RBCs also show a similar OF response to dicarboxylic acids in this experiment.

Highlights

  • Mammalian red blood cells (RBCs) have been used as a valuable tool, as an experimental model for the cell membranes composed of a phospholipid bilayer, as RBCs possess a basic structure consisting of a cell membrane without a nucleus or complex metabolic system in the plasma

  • The RBCs of various mammalian species were used in those experiments, it has not been considered if the differences in Osmotic fragility (OF) response are based on inter-species differences in RBC membrane characteristics

  • The results revealed that most monocarboxylic acids possessing straight hydrocarbon chains did not affect OF in cattle RBCs, except that the application of

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Summary

INTRODUCTION

Mammalian red blood cells (RBCs) have been used as a valuable tool, as an experimental model for the cell membranes composed of a phospholipid bilayer, as RBCs possess a basic structure consisting of a cell membrane without a nucleus or complex metabolic system in the plasma. The reason for the common OF response to dicarboxylic acids is assumed to be that the membrane resistance to osmotic pressure is strengthened by interaction between the conformation of the dicarboxylic acids and the structure of phospholipids close to the water/lipid layer, decreasing the OF in the RBCs in various animal species [20,30,31] The objectives of this experiment were to clarify the effects of the application of monocarboxylic and dicarboxylic acids on OF in cattle RBCs and to compare the OF responses to these chemicals with those in rat, guinea pig and sheep RBCs. The results of the present study allow us to the data on the OF response in cattle RBCs to the previous data obtained using the RBCs of different three mammalian species. We can confirm the validity about our hypotheses or action model, that we already proposed, for both carboxylic acids on the RBC membrane in various animal species

MATERIALS AND METHODS
Experimental procedure
RESULTS
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