Influence of mouse age and erythrocyte age on glutathione metabolism

Abraham, E.C.; Taylor, John Fuller; Lang, Calvin A.

doi:10.1042/bj1740819

Cited by 113 publications

(29 citation statements)

References 19 publications

(12 reference statements)

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“…As mentioned above, the total glutathione and 2,3-bisphosphoglycerate levels do not change during the aging of these cells. These results are contradictory to earlier reports of declines in both of these metabolites in "aged" human erythrocytes isolated by density gradient centrifugation (18)(19)(20)(21)(22). These discrepancies may be related to species differences or to the inadequacies of the density gradient separation methods.…”

Section: Discussioncontrasting

confidence: 56%

Senescent erythrocytes: isolation of in vivo aged cells and their biochemical characteristics.

Suzuki

Dale

1988

Proc. Natl. Acad. Sci. U.S.A.

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Rabbit erythrocytes were covalently labeled with biotin and then infused into the donor animial. Up to 60 days after infusion, the biotinylated cells were selectively isolated by their affinity for avidin. These aged erythrocytes, which are within 10 days of death, were analyzed for several biochemical parameters. The 2,3-bisphosphoglycerate and glutathione levels of these cells were constant with age; however, the adenosine 5'-triphosphate concentrations increased approximately 75% as the cells approached the end of their life span. The activities of several glycolytic enzymes did not change with age.The mechanisms of erythrocyte senescence are poorly understood (1). Clearly, erythrocytes have a well-defined life span; however, the study of the aging processes in these cells has been hindered by the inability to definitively isolate senescent erythrocytes. A myriad of publications have reported age-related differences in various biochemical parameters of erythrocytes (for a review, see ref. 1). However, the majority of these are based on the separation of cells over density gradients with the underlying assumption that the most dense fractions represent the old cells, an assumption that has been questioned (2,3).Two methods that are independent of the physical separation techniques outlined above have been reported for the isolation of senescent erythroctyes. Allison and Burn (4) transfused blood group 0 erythrocytes into normal blood group A individuals. At subsequent time points, the 0 cells were recovered after agglutination of the host A erythrocytes. Unfortunately, the technique was hindered by technical difficulties. More recently, Ganzoni et al. (5) Preparation of Young Erythrocyte Cohorts. In some experiments, young erythrocyte populations were used for biotinylation and replacement into donor rabbits. The advantage of using a young cohort of cells is that a larger percentage of the cells will still be circulating 50-60 days after replacement and therefore will allow recovery of greater numbers of senescent cells. Two procedures were used to produce young cell populations-phenylhydrazine treatment and repeated phlebotomy. Phenylhydrazine hydrochloride (25 mg) was subcutaneously injected into a rabbit on days 0, 2, and 3. On day 13, 50 ml of blood was collected in heparin for biotinylation. The reticulocyte count for these preparations averaged more than 50% on day 8. For the repeated phlebotomy procedure, 50-60 ml of blood was drawn from the rabbit on day 0, 2, and 4. The sample for biotinylation was taken on day 14. To protect against iron deficiency during repeated phlebotomy, each rabbit received 50 mg of iron-dextran intramuscularly on days -7, -5, -3, 0, 2, and 4. Preparation and Infusion of Biotinylated Erythrocytes.Rabbit erythrocytes were washed, labeled with ['4C]cyanate, treated with NHS-biotin, and infused into the donor rabbit exactly as described (7). The average level of biotinylation was approximately 25,000 biotin molecules per erythrocyte.Avidin-Coated Polystyrene Beads. The av...

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Section: Discussioncontrasting

confidence: 56%

Senescent erythrocytes: isolation of in vivo aged cells and their biochemical characteristics.

Suzuki

Dale

1988

Proc. Natl. Acad. Sci. U.S.A.

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“…Radiolabeled ribavirin transport rate was normalized to micrograms of protein (using the bicinchoninic acid assay and bovine serum albumin protein concentration standard curve) or cell count (hematocytometer). Intracellular ribavirin (or metabolite) concentrations were estimated by using an intracellular erythrocyte volume of 90 fl ( 3 ) in humans (Bessman and Johnson, 1975) and 48 fl (Abraham et al, 1978) in mice.…”

Section: Methodsmentioning

confidence: 99%

“…Healthy human erythrocytes have a fixed life span of approximately 120 days (Landaw, 1991), whereas in mice this life span is approximately 104 days (Abraham et al, 1978). Multiple dosing of ribavirin in monkeys has shown that the erythrocyte degradation was increased upon multiple dosing of ribavirin (Canonico et al, 1984).…”

Section: Downloaded Frommentioning

confidence: 99%

The Role of the Equilibrative Nucleoside Transporter 1 (ENT1) in Transport and Metabolism of Ribavirin by Human and Wild-Type or Ent1(-/-) Mouse Erythrocytes

Endres¹,

Moss²,

Ke³

et al. 2009

J Pharmacol Exp Ther

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The polar nucleoside drug ribavirin is front-line treatment for chronic hepatitis C virus infection. The human equilibrative nucleoside transporter (ENT) 1 transports ribavirin into erythrocytes where it is phosphorylated. These phosphorylated metabolites accumulate in the erythrocytes and produce doselimiting hemolytic anemia. Here, we examined the in vitro and ex vivo transport and metabolism of ribavirin by erythrocytes isolated from humans and Ent1-null mice. Ribavirin (2.4 M) uptake was significantly higher (1044 Ϯ 255 amol/g/10 s) into erythrocytes from Ent1(ϩ/ϩ) mice compared with that from Ent1(Ϫ/Ϫ) mice (76.48 Ϯ 11.20 amol/g/10 s). Our results showed a saturable (K m of 382 Ϯ 75.1 M) transport of [ 3 H]ribavirin into erythrocytes from Ent1(ϩ/ϩ) mice. We found that ribavirin concentration rapidly (within 60 s) reached equilibrium in erythrocytes using a time course of [ 3 H]ribavirin transport (2.5 M) and metabolism in mouse and human erythrocytes for 8 h. However, total radioactivity of ribavirin was predominantly attributed to the phosphorylated metabolites ribavirin monophosphate and ribavirin triphosphate. Our findings allow us to estimate ribavirin transport, diffusion, and metabolic clearance and to predict in vivo accumulation of ribavirin phosphates in erythrocytes of both mice and humans. Our modeling of ribavirin in erythrocytes on long-term administration of ribavirin suggests that the accumulation of ribavirin inside the cells is dependent on ENT1/Ent1 transport and the rates of intracellular phosphorylation and the degradation of the phosphorylated metabolites. We predict that Ent1(ϩ/ϩ) and Ent1(Ϫ/Ϫ) mice will serve as excellent models to investigate the contribution of Ent1 to the pharmacokinetics and toxicity of ribavirin in vivo.

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“…There is extensive literature on the metabolic changes that occur during cell aging (1,11,39,55,56,75,77,98,107). Changes have been reported for the antioxidant enzymes SOD1, catalase, and GPx (35).…”

Section: Rbc Antioxidant Systemmentioning

confidence: 99%

“…There are, however, uncertainties regarding many of the specific changes that occur. A number of studies indicate changes in glutathione metabolism as a function of cellular age including changes in enzymes involved in the hexose monophosphate shunt (1,35,39,40). This includes changes in glutathione, glutathione reductase, and glucose-6-phosphate dehydrogenase, which produces the NADPH required for glutathione reduction.…”

Section: Rbc Antioxidant Systemmentioning

confidence: 99%

Hemoglobin Redox Reactions and Red Blood Cell Aging

Rifkind

Nagababu

2013

Antioxidants & Redox Signaling

133

112

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Significance: The physiological mechanism(s) for recognition and removal of red blood cells (RBCs) from circulation after 120 days of its lifespan is not fully understood. Many of the processes thought to be associated with the removal of RBCs involve oxidative stress. We have focused on hemoglobin (Hb) redox reactions, which is the major source of RBC oxidative stress. Recent Advances: The importance of Hb redox reactions have been shown to originate in large parts from the continuous slow autoxidation of Hb producing superoxide and its dramatic increase under hypoxic conditions. In addition, oxidative stress has been shown to be associated with redox reactions that originate from Hb reactions with nitrite and nitric oxide (NO) and the resultant formation of highly toxic peroxynitrite when NO reacts with superoxide released during Hb autoxidation. Critical Issues: The interaction of Hb, particularly under hypoxic conditions with band 3 of the RBC membrane is critical for the generating the RBC membrane changes that trigger the removal of cells from circulation. These changes include exposure of antigenic sites, increased calcium leakage into the RBC, and the resultant leakage of potassium out of the RBC causing cell shrinkage and impaired deformability. Future Directions: The need to understand the oxidative damage to specific membrane proteins that result from redox reactions occurring when Hb is bound to the membrane. Proteomic studies that can pinpoint the specific proteins damaged under different conditions will help elucidate the cellular aging processes that result in cells being removed from circulation. Antioxid. Redox Signal. 18, 2274-2283.

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Influence of mouse age and erythrocyte age on glutathione metabolism

Cited by 113 publications

References 19 publications

Senescent erythrocytes: isolation of in vivo aged cells and their biochemical characteristics.

Senescent erythrocytes: isolation of in vivo aged cells and their biochemical characteristics.

The Role of the Equilibrative Nucleoside Transporter 1 (ENT1) in Transport and Metabolism of Ribavirin by Human and Wild-Type or Ent1(-/-) Mouse Erythrocytes

Hemoglobin Redox Reactions and Red Blood Cell Aging

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