In vitro enhancement of ATP in human erythrocytes from a healthy subject and two patients with thalassemia and hemoglobinopathy.

Abstract

Objective: To examine whether the supplementation of inosine augments ATP in vitro in human erythrocytes incubated in saline. Methods : Peripheral blood was drawn from each of three subjects, i.e. one healthy male and two males with thalassemia and hemoglobinopathy. After washing the erythrocytes in saline, they were suspended in saline to which inosine was added to final concentrations of 0, 0.5 and 2.5 mM. The suspension was incubated at 37 °C for 1 or 3 hours, and 0.5 ml ice cold 8% perchloric acid was added to the 0.5 ml erythrocyte-containing solution. After removing precipitates and perchloric acid, the supernatant was submitted to HPLC for the measurement of ATP. Results : Since the blood samples of the two subjects with thalassemia and hemoglobinopathy were transported from the clinics to the laboratory, ATP in the blood decreased considerably during the transportation. However, the reduction of ATP with time was observed in the erythrocytes in saline obtained from each of the three subjects during the incubation from 1 hour to 3 hours. In addition, dosedependent suppression of the decrease of ATP with inosine was observed in all the three cases at both 1 hour and 3 hour incubation times. Conclusions : Incubation of erythrocytes from a healthy subject and two thalassemia /hemoglobinopathy patients in saline at 37 °C resulted in time-dependent decreases of ATP. Supplementation of inosine to the solutions resulted in the suppression of the decreases of ATP in a dose-dependent manner. Background ATP(adenosine triphosphate)is the most important compound for storing energy in living organisms and supplying it when needed. It is considered that ATP reduction is related to the pathology of various diseases. An example is the hemolytic anemia caused by glycolytic enzyme deficiencies such as hexokinase deficiency1), pyruvate kinase deficiency2), glucose phosphate isomerase deficiency3), phosphofructokinase deficiency4)and phosphoglycerate kinase deficiency5). Since mature erythrocytes lack mitochondria, they are heavily dependent on the anaerobic generation of ATP during glycolysis for nearly all of their 受付:2018年4月2日,受理:2018年4月25日 1) Tsukuba International Clinical Pharmacology Clinic, Tsukuba, Japan 2) StaGen Co. Ltd, Tokyo, Japan 3) P-One Clinic, Keikokai Medical Corporation, Hachioji, Tokyo, Japan 4) Institute of Rheumatology, Tokyo Women's Medical University, Tokyo, Japan 5) Laboratory of Biomedical and Analytical Sciences, Faculty of Pharma-Science, Teikyo University, Tokyo, Japan 6) Department of Transfusion Medicine and Cell Processing, Tokyo Women's Medical University, Tokyo, Japan