In Vitro Inhibition of Blood Clycolysis with Wat

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1) In vitro inhibition of blood glycolysis by dilution with water has been reported by SAITO (1958) 1) This inhibiting effect of water is possibly due to dilution of enzyme and co-factor concentrations to the point where phosphorylation can not readily occur. However, the dilution of blood with physiologic saline solution causes very weak inhibition, but a strong one is caused by mechanical hemolysis without dilution with water (Fig.1, 2) 2)To investigate the inhibition of glycolysis by mechanical hemolysis of erythrocytes, some of glycolytic intermediates and nucleotides were enzymatically analyzed. ATPase activity was measured by the method of Bailey. Fig.3, 4 show a marked decrease in ATP and NAD levels and a high activity of ATPase in erythrocyte hemolysates. There was a high significant correlation between the ATP levels and the ATPase activity. The low levels of NAD appear to be a result of the fact that the decreased amount of pyruvate are present to be converted into lactate, because after incubation at 37°C for 1 hour, glucose converted into pyruvate only in the hemolysate (Fig.4). On the contrary, almost all of glucose used was converted into lactate in non hemolytic erythrocytes. 3) The studies of Murphy (1960) 7) demonstrated that approximately 90 per cent of all glucose metabolized under the normal conditions passes via the Embdem-Meyerhof pathway, an anaerobic pathway, ATP and NAD serving as an essential accelerator and cofactor in this route. The data presented in this study (Fig. 5) also indicate that addition of ATP and NAD to erythrocyte hemolysates in which glycolysis are almost retarded, the rate of glucose utilization is markedly increased and almost same as compared with the non hemolytic erythrocytes.

In Vitro Inhibition of Blood Clycolysis with Wat

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