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Cerebral metabolism of glucose, one of the determinants of tissue ATP level, is crucial for central nervous system function. The activity of P-type pumps, namely Na+, K+ - ATPase, Ca+2 - ATPase and Mg+2 - ATPase were examined in brain synaptosomes of 5 - day, 3 - month and 18 - month - old rats to determine if changes in enzyme activity related to aging are potentially associated with alterations in glucose homeostasis. Activities of all the ATPases studied in isolated brain synaptosomes were expressed in µmol of Pi liberated from ATP by 1 mg of synaptosome protein during one hour. Serum glucose concentration was measured by the glucose oxidase method and insulin level was estimated by the RIA. Our results demonstrate that 18 - month - old rats are characterized by hyperglycemia and hyperinsulinemia. Their serum glucose concentration was significantly increased approx. 62.3% and 135.8 % as compared to 3 - month - old rats and 5 - day, newborn rats, respectively. An enormous increase in serum insulin concentration in the old, hyperglycemic rats was observed concomitantly. As a result of these changes the insulin - to - glucose ratio in the old rats was greatly increased approx. (270% and 230%) compared to young, mature and newborn rats. Hyperglycemia and hyperinsulinemia occurring in the old rats, had a different impact on activities of the ATPases tested. Our results have revealed that Na+, K+ - ATPase activity remains almost unchanged with age, the activity of Ca+2 - ATPase decreases, whereas that of Mg+2 - ATPase increases significantly in old, insulin resistant rats. In conclusion it seems that changes in activity of different P - type pumps may differ with aging and that adaptation of specific ATPases to internal environment alterations is not identical.
Torlińska T., Rutkowska D., Hryniewiecki T., Paluszak J.: In vivo effect of 2-deoxy-D-glucose on adenine nucleotide levels in the liver and skeletal muscle. Acta Physiol. Pol. 1990, 41 (7): 75-83. The present report indicates that 2-deoxy-D-glucose (2-DG) at a single dose causing reduction of Т„ has no influence on liver and skeletal muscle content of ATP, ADP and AMP, the ATP/ADP ratio, energy charge potential (ECP) and total adenine nucleotides (TAN). After administration of 2-DG for 3) successive days, the level of ATP, ATP/ADP ratio, the values of ECP and TAN are decreased both in the liver nad skeletal muscle. However, 72 hours after the last injection of 2-DG adenine nucleotide contents returned to the values observed in control group, indicating that the in vivo effect of this glucose analogue is fully reversible.
Torlińska T., Ożegowski S., Paluszak J., and Hryniewiecki T.: In vivo effect of 2-deoxy-D-glucose on glucose-6-phosphate dehydrogenase activity in the cytosol of liver, heart and skeletal muscle of rats. Acta Physiol. Pol. 1990, 41 (6): 137-143. 2-deoxy-D-glucose (2-DG), the unmetabolizable analogue of glucose induces a series of metabolic, hormonal and behavioral responses, causing cellular glucoprivation. According to in vitro studies, 2-DG inhibits phosphofructokinase in cultured human cells. The present investigations deal with changes in the cytosolic glucocse-6-phosphate dehydrogenase activity following in vivo 2-DG administration. A single dose of 2-DG (600 mg/kg) has no influence on the activity of glucose-6-phosphate dehydrogenase in the cytosol of liver, heart and skeletal muscle of the rat. The concommitant increase in serum glucose, lactate and FFA concentrations observed in the study indicates indirectly a stimulation of adrenergic system. After three days of successive administration of 2-DG to rats, dehydrogenase activity decreased in the liver by approx 57% and in the skeletal muscle by approx 82% in comparison with control animals. Moreover the in vivo effect of 2-DG was found to be fully reversible, probably when the total amount of the inhibitor was excreted.
Cerebral metabolism of glucose, one of the determinants of tissue ATP level, is crucial for the CNS function. The activity of P-type pumps: Na+, K+-ATPase, Ca+2-ATPase and Mg+2-ATPase were examined in rat brain synaptosomes to determine if changes in the enzyme activity related to aging are potentially associated with alterations in glucose homeostasis. Male Wistar rats (newborn, 3- and 18-month-old) were sacrificed by decapitation and synaptic plasma membranes were isolated from brains. In vivo study demonstrated that 18-month-old rats were characterized by hyperglycemia, hyperinsulinemia and increased total antyoxidative status (TAS) level. These conditions had a different impact on activities of the ATPases tested in vivo: only the activity of Ca+2-ATPase decreased whereas that of Mg+2-ATPase increased significantly. In vitro experiments, prior incubation of isolated synaptosomes with glucose of concentrations corresponding to normoglycemia in vivo (4.5 - 6.5 mM), stimulated Ca+2-ATPase activity, whereas higher glucose concentrations (10.0 - 12.5 mM) inhibited significantly the enzyme activity. The most sensitive to hyperglycemia appeared Na+, K+-ATPase in old rats synaptosomes with the progressive decline starting at 6.5 mM glucose. The activity of Mg+2-ATPase was not inhibited in vitro even at high glucose concentrations that may explain the increased in vivo, activity of this enzyme in old, hyperglycemic rats.
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