In vitro studies on the entry of polyamines into normal red blood cells

Moulinoux, Jacques-Philippe; Calve, Michèle Le; Quemener, V.; Quash, G.

doi:10.1016/0300-9084(84)90022-1

Cited by 49 publications

(19 citation statements)

References 5 publications

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“…If the NO donor is transported into RBC, then the NO consumption rate by the RBC measured by the competition experiment would be higher than the true value. In this study, we used spermine NONOate as an NO donor, which does not enter RBC at an appreciable rate (22). The amount of polyamines that might enter RBC within the time scale of our experiment is too small to affect our results.…”

Section: Discussionmentioning

confidence: 99%

Erythrocytes Possess an Intrinsic Barrier to Nitric Oxide Consumption

Vaughn

Huang

et al. 2000

Journal of Biological Chemistry

211

196

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It has been reported that free hemoglobin (Hb) reacts with NO at an extremely high rate (K Hb ϳ10 M؊1 s ؊1 ) and that the red blood cell (RBC) membrane is highly permeable to NO. RBCs, however, react with NO 500 -1000 times slower. This reduction of NO reaction rate by RBCs has been attributed to the extracellular diffusion limitation. To test whether additional limitations are also important, we designed a competition test, which allows the extracellular diffusion limitation to be distinguished from transmembrane or intracellular resistance. This test exploited the competition between free Hb and RBCs for NO generated in a homogenous phase by an NO donor. If the extracellular diffusion resistance is negligible, then the results would follow a kinetic model that assumes homogenous reaction without extracellular diffusion limitation. In this case, the measured effective reaction rate constant, K RBC , would remain invariant of the hematocrit, extracellular-free Hb concentration, and NO donor concentration. Results show that the K RBC approaches a constant only when the hematocrit is greater than 10%, suggesting that at higher hematocrit, the extracellular diffusion resistance is negligible. Under such a condition, the NO consumption by RBCs is still 500 -1000 times slower than that by free Hb. This result suggests that intrinsic RBC factors, such as transmembrane diffusion limitation or intracellular mechanisms, exist to reduce the NO consumption by RBCs.Despite the well documented importance of nitric oxide, the transfer of NO from the producing cell to the target is poorly understood, because the free radical NO can be degraded in a variety of reactions. In particular, NO reacts with deoxy-and oxyhemoglobin (deoxyHb and oxyHb, respectively) 1 at a very high rate (1, 2) to form nitrosyl Hb (HbNO) and met hemoglobin (metHb), respectively. If Hb in the RBC behaved like Hb in dilute solution, the half-life of NO in the blood (which contains about 12-15 mM heme) would be only about 1 s. For such a rapid reaction, it would seem likely that a large portion of NO produced from the endothelium would be scavenged by the blood.Indeed, in vivo and in vitro evidence suggests that free Hb is an effective NO scavenger that can deplete NO. For example, infusion of free Hb solution into experimental animals or human subjects results in hypertension (3, 4), most likely due to the reaction of NO with oxyHb in the circulation (3). Modeling analyses (5-7) also showed that if endothelium-produced NO reacted as rapidly with blood as it does with free Hb, the NO concentrations in vascular smooth muscle would be too low to activate soluble guanylate cyclase, the primary target of NO. Mathematical modeling based on diffusion theory and in vitro measurements of kinetic constants (5-8) have confirmed that Hb could effectively scavenge endothelial produced NO and mitigate its effect. Because 3-10 M free Hb can abolish NOmediated vasodilation in vitro (4, 9), it is unclear how NO can exercise its vasoregulatory function with 12-15 mM Hb c...

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Section: Discussionmentioning

confidence: 99%

Erythrocytes Possess an Intrinsic Barrier to Nitric Oxide Consumption

Vaughn

Huang

et al. 2000

Journal of Biological Chemistry

211

196

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“…After release from brain tissue, polyamines are mainly transported in the blood by erythrocytes. 30,31 In the presence of serum, the affinity of red blood cells for spermidine is 30-fold greater than that for other polyamines, 30 which may explain the difficulties in the measurement of spermine and putrescine. …”

Section: Discussionmentioning

confidence: 99%

Spermidine: A Predictor for Neurological Outcome and Infarct Size in Focal Cerebral Ischemia?

Els

Bruckmann²,

Röhn³

et al. 2001

Stroke

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Background and Purpose-Polyamines are mainly restricted to the intracellular space. During focal cerebral ischemia, polyamines are released from the intracellular compartment. Experimental studies have implicated a marked elevation in brain tissue and blood. The aim of our study was to investigate whether the elevation of polyamines in the blood of patients with focal cerebral ischemia correlates with the clinical outcome and the infarct volume. Methods-Polyamines were measured in 16 patients with focal cerebral ischemia and in 8 healthy control subjects. Blood samples for polyamine measurement were taken at admission and at fixed time points for the next 28 days. Polyamines were analyzed in red blood cells by a high-pressure liquid chromatography system. Clinical findings were recorded with the NIH Stroke Scale score. Volume of infarction was analyzed from cranial CT at admission and on days 4 to 6 after ischemia. Results-A significant increase of the spermidine level in the peripheral blood could be observed in all patients with focal cerebral ischemia as compared with control subjects (

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“…The homologous bis-a-<o-dimethylaminoalkanes were obtained from the respective a-to-diamines by permethylation with formaldehyde/sodium borohydride [20]. The following com pounds were prepared (as dihydrochlorides) in a chromatographically pure form: N,N'-tetramethyl-l,2-ethanediamine (1,2-TME), N,N'-tetramethyl-l, , N,N-tetramethyl-l,4-butanediamine (1,, N,N'-tetramethyl-1,5-pentanediamine (1,, N,N'-tetramethyl-l,6-hexanediamine , N,N'-tctramethyl-l,7-hcptanediamine (1,7-TMH). 2-fluoroputrescine (FP) was synthesized according to Saito et al [21], 2,2-Difluoroputrescine (DFP) was prepared as described elsewhere [22].…”

Section: Methodsmentioning

confidence: 99%

“…In a previous study, we demonstrated the existence of polyamine-specific acceptor sites at the cell surface of human erythrocytes [1] and reported that red blood cell affinity for spermidine (Spd; Km 3.6 \iM) was signifi cantly higher than that observed for putrescine (Put; Km 125 pAf). Recently, we showed that erythrocyte plasma membrane proteins, particularly band 3 proteins, are modified in cancer-grafted mice.…”

Section: Introductionmentioning

confidence: 99%

Mechanism of Spermidine Uptake in Cultured Mammalian Cells and Its Inhibition by Some Polyamine Analogues

Khan

Quemener

Seiler³

et al. 1990

Pathobiology

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Transport pathways for spermidine (Spd) were characterized in mammalian cells in culture of different origin, i.e. L 1210, P 388, C 6, U 251, Balb/c 3T3 normal and transformed by virus SV40 (SV40/3T3). The kinetic constants (K_m and V_max) for ¹⁴C-Spd uptake were found to be different in these cells. Spd uptake was inhibited by spermine and putrescine in all cells. Preloading of these cells with system A and other amino acids, including ornithine, usually did not affect Spd uptake, except in L 1210 and C 6 cells, where Spd uptake was accelerated by 2-aminoisobutyric acid, demonstrating that in these two cell lines the polyamines share the system A pathway. Iso-osmotic replacement of Na⁺ by choline chloride in the assay medium resulted in a decrease in Spd uptake which suggests that Spd uptake is Na⁺ activated. In all cells, Spd uptake was inhibited by gramicidin and the Ca²⁺ ionophore A 23187. The degree of inhibition varied among the cells. Valinomycin (K⁺ ionophore) inhibited Spd uptake by C 6, P 388, Balb/c 3T3 and SV40/3T3 but not by L 1210 and U 251 cells. Treatment with N-ethylmaleimide or p-chloromercuribenzene sulfonate abolished the Spd uptake by U 251 and P 388 cells, while the same treatment to L 1210, C 6, Balb/c 3T3 and SV40/3T3 cells did not affect appreciably the uptake process. Some newly synthesized polyamine analogues inhibited the Spd uptake of all cells.

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In vitro studies on the entry of polyamines into normal red blood cells

Cited by 49 publications

References 5 publications

Erythrocytes Possess an Intrinsic Barrier to Nitric Oxide Consumption

Erythrocytes Possess an Intrinsic Barrier to Nitric Oxide Consumption

Spermidine: A Predictor for Neurological Outcome and Infarct Size in Focal Cerebral Ischemia?

Mechanism of Spermidine Uptake in Cultured Mammalian Cells and Its Inhibition by Some Polyamine Analogues

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