Injections of recombinant human erythropoietin increases lactate influx into erythrocytes

2004 ◽  
Vol 97 (1) ◽  
pp. 326-332 ◽  
Author(s):  
Philippe Connes ◽  
Corinne Caillaud ◽  
Jacques Mercier ◽  
Didier Bouix ◽  
Jean François Casties
Keyword(s):  
Placebo Group ◽  
Recombinant Human Erythropoietin ◽  
Lactate Concentration ◽  
Band 3 ◽  
Monocarboxylate Transporter ◽  
Maximal Velocity ◽  
Lactate Transport ◽  
Human Erythropoietin ◽  
Before And After ◽  
Lactate Uptake

Previous studies showed that erythropoietin not only increases erythrocyte production but is also essential in both the synthesis and the good functioning of several erythrocyte membrane proteins, including band 3. It is still unknown whether anion and/or H+ fluxes are modified by erythropoietin. This study aimed to evaluate the effect of recombinant human erythropoietin (rHuEPO) injections on lactate transport into erythrocytes via band 3 and H+-monocarboxylate transporter MCT-1, two proteins involved in lactate exchange. Nine athletes received subcutaneous rHuEPO (50 U/kg body mass 3 times a week for 4 wk), and seven athletes received a saline solution (placebo group). All subjects were also supplemented with oral iron and vitamins B9 and B12. Lactate transport into erythrocytes was studied before and after the rHuEPO treatment at different lactate concentrations (1.6, 8.1, 41, and 81.1 mM). After treatment, MCT-1 lactate uptake was increased at 1.6, 41 ( P < 0.01), and 81.1 mM lactate concentration ( P < 0.001) although lactate uptake via band 3 and nonionic diffusion were unchanged. MCT-1 maximal velocity increased in the rHuEPO group ( P < 0.05), reaching higher values than in the placebo group ( P < 0.05) after treatment. Our results show that rHuEPO injections increased MCT-1 lactate influx at low and high lactate concentrations. The increase in MCT-1 maximal velocity suggests that rHuEPO may stimulate MCT-1 synthesis during erythrocyte formation in bone marrow.

Muscle contraction increases lactate transport while reducing sarcolemmal MCT4, but not MCT1

2002 ◽  
Vol 282 (5) ◽  
pp. E1062-E1069 ◽  
Author(s):  
Mio Tonouchi ◽  
Hideo Hatta ◽  
Arend Bonen
Keyword(s):  
Plasma Membrane ◽  
Muscle Contraction ◽  
Soleus Muscle ◽  
Muscle Glycogen ◽  
Lactate Concentration ◽  
Monocarboxylate Transporter ◽  
Intrinsic Activity ◽  
Lactate Transport ◽  
Lactate Uptake ◽  
Sarcolemmal Vesicles

Rates of lactate uptake into giant sarcolemmal vesicles were determined in vesicles collected from rat muscles at rest and immediately after 10 min of intense muscle contraction. This contraction period reduced muscle glycogen rapidly by 37–82% in all muscles examined ( P < 0.05) except the soleus muscle (no change P > 0.05). At an external lactate concentration of 1 mM lactate, uptake into giant sarcolemmal vesicles was not altered ( P > 0.05), whereas at an external lactate concentration of 20 mM, the rate of lactate uptake was increased by 64% ( P < 0.05). Concomitantly, the plasma membrane content of monocarboxylate transporter (MCT)1 was reduced slightly (−10%, P < 0.05), and the plasma membrane content of MCT4 was reduced further (−25%, P < 0.05). In additional studies, the 10-min contraction period increased the plasma membrane GLUT4 ( P < 0.05) while again reducing MCT4 (−20%, P < 0.05) but not MCT1 ( P > 0.05). These studies have shown that intense muscle contraction can increase the initial rates of lactate uptake, but only when the external lactate concentrations are high (20 mM). We speculate that muscle contraction increases the intrinsic activity of the plasma membrane MCTs, because the increase in lactate uptake occurred while plasma membrane MCT4 was decreased and plasma membrane MCT1 was reduced only minimally, or not at all.

Recombinant Human Erythropoietin Stimulates Erythropoiesis and Reduces Erythrocyte Transfusions in Very Low Birth Weight Preterm Infants

PEDIATRICS ◽  
1995 ◽  
Vol 95 (1) ◽  
pp. 1-8
Author(s):  
Kevin M. Shannon ◽  
Julian F. Keith ◽  
William C. Mentzer ◽  
Richard A. Ehrenkranz ◽  
Mark S. Brown ◽  
...  
Keyword(s):  
Placebo Group ◽  
Birth Weight ◽  
Low Birth Weight ◽  
Preterm Infants ◽  
Recombinant Human Erythropoietin ◽  
Very Low Birth Weight ◽  
Group Versus ◽  
Weekly Dose ◽  
Human Erythropoietin ◽  
Transfusion Requirements

Design and methods. We hypothesized that treatment with recombinant human erythropoietin (r-HuEPO) would stimulate erythropoiesis and would thereby reduce the need for erythrocyte transfusions in preterm infants. We treated 157 preterm infants born at 26.9 ± 1.6 weeks of gestation who weighed 924 ± 183 g at birth with either subcutaneous r-HuEPO (100 U/kg/d, 5 days per week) or placebo for 6 weeks in a randomized, double-blind, controlled clinical trial. All patients received oral iron and were managed according to uniform conservative transfusion guidelines. Results. Treatment with r-HuEPO was associated with fewer erythrocyte transfusions (1.1 ± 1.5 per infant in the r-HuEPO group versus 1.6 ± 1.7 per infant in the placebo group; P = .046) and with a reduction in the volume of packed erythrocytes transfused (16.5 ± 23.0 mL versus 23.9 ± 25.7 mL per infant; P = .023). Overall, 43% of the infants in the r-HuEPO group and 31% of placebo-treated infants were transfusion-free during the study (P = .18). The volume of blood removed for laboratory tests and the need for respiratory support at the start of treatment had major effects on transfusion requirements independent of r-HuEPO. Reticulocyte counts were higher during treatment in the r-HuEPO group (P = .0001), and r-HuEPO-treated infants had higher hematocrit values at the end of the study (32% versus 27.3% in the placebo group; P = .0001). We found no differences in the incidence of major complications of prematurity between the treatment groups. Conclusion. We conclude that treatment with r-HuEPO at a weekly dose of 500 U/kg stimulates erythropoiesis, moderates the course of anemia, is associated with a reduction in erythrocyte transfusions, and appears safe in very low birth weight preterm infants who are receiving iron supplements. Conservative transfusion criteria, minimization of phlebotomy losses, and treatment with r-HuEPO are complementary strategies to reduce erythrocyte transfusions in these infants.

Quality of Life Improvements in Capd Patients Treated with Subcutaneously Administered Erythropoietin for Anemia

1992 ◽  
Vol 12 (1) ◽  
pp. 40-42 ◽  
Author(s):  
Juliet Auer ◽  
Gail Simon ◽  
Judith Stevens ◽  
Paul Griffiths ◽  
Deborah Howarth ◽  
...  
Keyword(s):  
Social Life ◽  
Recombinant Human Erythropoietin ◽  
Nottingham Health Profile ◽  
Hemodialysis Patients ◽  
Health Profile ◽  
Emotional Wellbeing ◽  
Household Tasks ◽  
Human Erythropoietin ◽  
Before And After

Twenty -two continuous ambulatory peritoneal dialysis (CAPD) patients, mean age 48 years, at 3 U.K. renal units were assessed with the Nottingham Health Profile (N HP) before and after treatment with recombinant human erythropoietin (r-HuEPO). Mean (SD) hemoglobin (Hb) at baseline was 7.5 (1.0) gIn/dl and 10.8 (1.5) gIn/dl at retest. There were significant improvements in energy (p<0.0001), social life (p<0.005), relationships at home (p<0.05) and leisure pursuits (p<0.05). Twelve patients, mean age 51 years, who had already completed more than 9 months on r-HuEPO treatment were reassessed to determine the changes sustained. Mean (SD) Hb at second retest was 12.8 (1.3) gm/dl. Improvement in energy continued to be significant, and emotional wellbeing showed further improvement. Problems with household tasks, which had not shown significant improvement at Test B, were now considerably reduced (p=0.016). The study showed far-reaching benefits similar to those reported in hemodialysis patients, in a population with a higher mean age and higher potential coexisting illness or disability than most reported hemodialysis studies.

Treatment of Anaemia in Haemodialysis Patients with Erythropoietin: Long-Term Effects on Exercise Capacity

1993 ◽  
Vol 84 (4) ◽  
pp. 441-447 ◽  
Author(s):  
Peter Báaráany ◽  
Ulla Freyschuss ◽  
Erna Pettersson ◽  
Jonas Bergström
Keyword(s):  
Oxygen Uptake ◽  
Blood Lactate ◽  
Exercise Capacity ◽  
Recombinant Human Erythropoietin ◽  
Maximal Exercise ◽  
Lactate Concentration ◽  
Blood Lactate Concentration ◽  
Human Erythropoietin ◽  
Hb Concentration ◽  
Maximal Exercise Capacity

1. The effects of correcting anaemia on exercise capacity were evaluated in 21 haemodialysis patients (aged 39 ± 12 years) before starting treatment with recombinant human erythropoietin (Hb concentration, 73 ± 10 g/l; total Hb, 59 ± 12% of expected), after correction of the anaemia to a Hb concentration of 108 ± 7 g/l and a total Hb 82 ± 10% of expected, and in 13 of the patients after 12 months on maintenance recombinant human erythropoietin treatment (Hb concentration 104 ± 14 g/l, total Hb 79 ± 17% of expected). Fifteen healthy subjects (aged 41 ± 9 years), who took no regular exercise, constituted the control group. Maximal exercise capacity was determined on a bicycle ergometer. Oxygen uptake, respiratory quotient, blood lactate concentration, heart rate and blood pressure were measured at rest and at maximal workload. 2. After 6 ± 3 months on recombinant human erythropoietin, maximal exercise capacity increased from 108 ± 27 W to 130 ± 36 W (P < 0.001) and the maximal oxygen uptake increased from 1.24 ± 0.39 litres/min to 1.50 ± 0.45 litres/min (P < 0.001). No significant changes in respiratory quotient (1.16 ± 0.13 versus 1.18 ± 0.13) and blood lactate concentration (4.0 ± 1.8 versus 3.6 ± 1.1 mmol/l) at maximal workload were observed, but the blood lactate concentration in the patients was significantly lower than that in the control subjects (6.7 ± 2.3 mmol/l, P < 0.01). After the correction of anaemia, the aerobic power was still 38% lower in the patients than in the control subjects and 17% lower than the reference values. 3. After 12 months on maintenance recombinant human erythropoietin treatment (17 ± 3 months from the start of the study), no further significant changes were observed in maximal exercise capacity (before start, 112 ± 31 W, 6 ± 3 months, 134 ± 42 W, 17 ± 3 months, 134 ± 50 W), maximal oxygen uptake (before start, 1.33 ± 0.45 litres/min; 6 ± 3 months, 1.59 ± 0.54 litres/min; 17 ± 3 months, 1.75 ± 0.78 litres/min) or blood lactate concentration (before start, 4.4 ± 1.9 mmol/l; 6 ± 3 months, 4.0 ± 1.0 mmol/l; 17 ± 3 months, 4.7 ± 2.0 mmol/l). 4. Thus, in haemodialysis patients the improvement in maximal aerobic power after the correction of anaemia persists without marked changes during long-term treatment with recombinant human erythropoietin. We did not observe any effects on exercise capacity that could be attributed to a spontaneous increase in physical activity after treatment of anaemia.

scholarly journals Characterization of the monocarboxylate transporter 1 expressed in Xenopus laevis oocytes by changes in cytosolic pH

1998 ◽  
Vol 333 (1) ◽  
pp. 167-174 ◽  
Author(s):  
Stefan BRÖER ◽  
Hans-Peter SCHNEIDER ◽  
Angelika BRÖER ◽  
Basim RAHMAN ◽  
Bernd HAMPRECHT ◽  
...  
Keyword(s):  
Xenopus Laevis ◽  
Intracellular Ph ◽  
Expression System ◽  
Lactate Concentration ◽  
Monocarboxylate Transporter ◽  
Kinetic Properties ◽  
Xenopus Laevis Oocytes ◽  
Lactate Transport ◽  
Monocarboxylate Transporter 1

Several laboratories have investigated monocarboxylate transport in a variety of cell types. The characterization of the cloned transporter isoforms in a suitable expression system is nevertheless still lacking. H+/monocarboxylate co-transport was therefore investigated in monocarboxylate transporter 1 (MCT1)-expressing Xenopus laevis oocytes by using pH-sensitive microelectrodes and [14C]lactate. Superfusion with lactate resulted in intracellular acidification of MCT1-expressing oocytes, but not in non-injected control oocytes. The basic kinetic properties of lactate transport in MCT1-expressing oocytes were determined by analysing the rates of intracellular pH changes under different conditions. The results were in agreement with the known properties of the transporter, with respect to both the dependence on the lactate concentration and the external pH value. Besides lactate, MCT1 mediated the reversible transport of a wide variety of monocarboxylic acids including pyruvate, d,l-3-hydroxybutyrate, acetoacetate, α-oxoisohexanoate and α-oxoisovalerate, but not of dicarboxylic and tricarboxylic acids. The inhibitor α-cyano-4-hydroxycinnamate bound strongly to the transporter without being translocated, but could be displaced by the addition of lactate. In addition to changes in the intracellular pH, lactate transport also induced deviations from the resting membrane potential.

scholarly journals Effect of 2-chloropropionate on initial lactate uptake by rat skeletal muscle sarcolemmal vesicles

1996 ◽  
Vol 81 (5) ◽  
pp. 1973-1977 ◽  
Author(s):  
P. Granier ◽  
H. Dubouchaud ◽  
N. Eydoux ◽  
J. Mercier ◽  
C. Préfaut
Keyword(s):  
Skeletal Muscle ◽  
Initial Rate ◽  
Lactate Concentration ◽  
Blood Lactate Concentration ◽  
Membrane Vesicles ◽  
Rat Skeletal Muscle ◽  
Lactate Transport ◽  
Sarcolemmal Membrane ◽  
Lactate Uptake ◽  
Sarcolemmal Vesicles

Granier, P., H. Dubouchaud, N. Eydoux, J. Mercier, and C. Préfaut. Effect of 2-chloropropionate on initial lactate uptake by rat skeletal muscle sarcolemmal vesicles. J. Appl. Physiol. 81(5): 1973–1977, 1996.—2-Chloropropionate (2-CP) is a halogenated monocarboxylic acid generally used to decrease blood lactate concentration in various metabolic states. To investigate whether it has an inhibitory effect on sarcolemmal lactate transport, we compared the initial rate of lactate transport in sarcolemmal membrane vesicles purified from 20 male Wistar rats with and without 2-CP. Transport by these vesicles was measured as uptake ofl-(+)-[U-14C]lactate under pH gradient-stimulated cisinhibition. The time courses of 1 mMl-(+)-lactate uptake into vesicles both with and without 10 mM 2-CP (l- ord-) displayed saturation kinetics. Lactate uptake values were lower with 10 mMl-2-CP and 10 mMd-2-CP in comparison to the control values. Both 10 mMl-2-CP and 10 mMd-2-CP significantly inhibited 1 mM l-(+)-lactate uptake (55.8 ± 9.1 and 53.5 ± 12.1%, respectively; P < 0.001), whereas a smaller inhibition was observed with a higher lactate concentration of 50 mM (40.2 ± 11.2 and 38.7 ± 12.4%; P < 0.001 and P < 0.05, respectively). However, a higher d-2-CP concentration (50 mM) increased the inhibition of pH-stimulated 1 mMl-(+)-lactate uptake (77.0 ± 9.4%; P < 0.001).d-2-CP had a trans-stimulation effect on the initial rate of lactate efflux of 1 mMl-(+)-lactate compared with baseline efflux (9.5 ± 0.8 vs. 5.1 ± 0.4 nmol ⋅ min−1 ⋅ mg protein−1; P < 0.05). 2-CP significantly inhibited the initial rate of lactate uptake in skeletal muscle sarcolemmal membrane vesicles. This result suggests that 2-CP is a nonstereoselective substrate of the lactate muscle carrier that impairs lactate transport.

First Report of a Prospective Randomised Controlled Trial of Recombinant Human Erythropoietin after Allogeneic Blood Stem Cell Transplantation.

Blood ◽  
2004 ◽  
Vol 104 (11) ◽  
pp. 1833-1833
Author(s):  
Anne Klink ◽  
Christoph Kasper ◽  
Sebastian Scholl ◽  
Lars-Olof Muegge ◽  
Kristina Schilling ◽  
...  
Keyword(s):  
Stem Cell ◽  
Placebo Group ◽  
Side Effects ◽  
Stem Cell Transplantation ◽  
Cell Transplantation ◽  
Recombinant Human Erythropoietin ◽  
Donor Group ◽  
Human Erythropoietin ◽  
Rbc Transfusion ◽  
Blood Stem Cell Transplantation

Abstract Recombinant human erythropoietin (rHuEPO) stimulates progenitor cells of erythropoiesis and increases erythrocyte production. This prospective, placebo-controlled, double-blind trial was designed to evaluate the use of erythropoietin versus red blood cell transfusions (RBC) to evade detrimental effects of anaemia during the first weeks after allogeneic peripheral blood stem cell transplantation (alloPBSCT). Between 4/2002 and 4/2004, 52 patients recieved either 10.000 units rHuEPO [Erypho(R)] 3 times a week subcutaneously or placebo, stratified according to AB0-incompatibility. Applications were started after stem cell transfusion (day +1) and lasted up until in mean day +39.5 [range: 29–50] or until haematocrit (HC) levels reached 0.38. Both groups underwent identical supportive care and RBC transfusion policy. Need for transfusion was defined by HC< 0.27. At time of transplantation, 18 (35%) patients were in remission, and 34 (65%) in advanced stage of their disease. Within the 22 female and 30 male patients (median age: 44.5 years [range:17–64]) 52 % (n=27) received for conditioning treatment a total body irradiation-based regimen, 4% (n=2) high dose treosulfan and cyclophosphamid, and 48 % (n=25) a dose reduced conditioning including fludarabine, busulfan/melphalan and anti-thymocyte globuline, respectively. Thirteen (25 %) patients received transplant from HLA-identical family donors, 26 (50%) from HLA-matched unrelated and 13 (25 %) from mis-matched alternative donors. The two study groups were well balanced regarding age, gender, disease status, graft characteristics, and conditioning regimens. Two patients were excluded from the study due to graft failure on day 21 and one patient refused further treatment on day 37. Eight side effects were observed in the rHuEPO group (3 x local haematoma, 3 x bone/muscle pain, 1 x dizziness, 1 x headache), whereas one was reported in the placebo group (1x headache). No serious adverse event related to study drug was noted. The mean numbers of RBC transfusions were 10.8 ± 5.5 [range: 4–18] with placebo and 9.9 ± 4.2 [range: 6–19] with rHuEPO up to day +50. These effects were identical in the unrelated donor group (n=24) with 11.4 ± 5.7 versus 10.4 ± 4.1, and in the HLA-identical family donor group (n=28) 10.8 ± 5.5 versus 9.4 ± 2.4. Interestingly, in the minor AB0-group (n=15) the rHuEPO arm needed 8.1 ± 3.4 as compared to 12.4 ± 4.2 RBC transfusion in the placebo arm. No differences were observed in the major AB0 group (n=12). Reticulocytes, by similar lab counts on day 0 in both groups, were significantly enhanced on day +30 in the rHuEPO arm. (62.6 ± 18.9 versus 29.3 ± 14.3; p < 0.001). Mean HC on day + 30 was 0.28 ± 0.03, on day + 50 0.33 ± 0.05 in the placebo arm, 0.31 ± 0.03 and 0.36 ± 0.04 in the rHuEPO arm, respectively. Overall survival of the total group after the first 100 days was 49/52 (94,2%).One (3.8%) patient died in placebo-group, and 2 (7.7%) patients in the rHuEPO-group. Further clinical parameters like engraftment, acute graft-versus-host disease (GvHD) and relapse rate were not different between the two treatment groups so far. After alloPBSCT rHuEPO can be given without initial clinical relevant side effects. We conclude that the reconstitution of erythropoiesis seems to be accelerated by early treatment with rHuEPO and reduces number of RBC-transfusions in AB0 minor incompatibility situation after alloPBSCT.

Molecular Pathways of Response to Recombinant Human Erythropoietin (rHuEpo) in Myelodysplastic patients Assayed with Macroarrays.

Blood ◽  
2004 ◽  
Vol 104 (11) ◽  
pp. 2367-2367
Author(s):  
Cortelezzi Agostino ◽  
Colombo Gualtiero ◽  
Pellegrini Caterina ◽  
Silvestris Ilaria ◽  
Bosari Silvano ◽  
...  
Keyword(s):  
Gene Expression ◽  
Bone Marrow ◽  
Recombinant Human Erythropoietin ◽  
Differentially Expressed ◽  
Monocarboxylate Transporter ◽  
Nuclear Antigen ◽  
Erythroid Cells ◽  
Down Regulation ◽  
Monocarboxylate Transporter 1 ◽  
Human Erythropoietin

Abstract Recently, among myelodysplastic syndromes with IPSS low/intermediate 1 risk classes, promising results have been obtained in the treatment of anemia by high dosage recombinant human erythropoietin (rHuEPO) treatment, in the cases with low endogenous erythropoietin (EPO). We have reproduced these clinical results in the patients treated at our Institution. Based on these premises, in order to evaluate the molecular basis of the rHuEPO response in MDS patients, we investigated the differential gene expression in the bone marrow erythroid cells expressing glycophorin (Gly+) of EPO-responders (ER) and EPO-non responders (ENR) MDS patients, using commercially available macroarrays (Clontech), comparing gene level of expression with a baseline represented by a pool of erythoid cells isolated from normal bone marrow donors. We analysed 4 ER and 4 ENR MDS patients (RA and RARS). BM erythroid cells, after Ficoll density gradient, were separated by positive selection using an immunomagnetic procedure (MACS, glycophorin isolation kit; Miltenyi Biotech, Auburn, CA). The purity of Gly+ cells after magnetic immunosorting was tested by flow-cytometry and was superior to 97%, in all the cases. RNA extraction, cDNA synthesis and macroarrays hybridization and analysis were performed following manufacturer instructions (Clontech Laboratories, Palo Alto, CA). The data were validated on 4 differentially expressed genes by real time RT-PCR TaqMan technology, obtaining results that were superimposable to the macroarrays ones. Using a cut off of an at least two fold difference in gene expression, we obtained a differential pattern of expression in ER and ENR patients, BM Gly+ erythroid cells, with respect to normal donors. In particular, ER patients presented the up-regulation of interleukin-1 beta, apolipoprotein E precursor, CREB2, high mobility group protein (HMG-I), transducin beta-1 subunit and the down regulation of protein kinase MLK-3, cAMP-dependent phosphodiesterase (PDE43), IL-5R-alpha, caspase-9. On the other hand, in ENR group, we found a down regulation of proliferating cyclic nuclear antigen (PCNA), B-myb, prothymosin alpha, phenylethanolamine N-methyltransferase, thymosin beta-10 and an up regulation of homeobox protein HOX-A5, monocarboxylate transporter 1, interleukin-6, deoxyribonuclease II, sterol regulatory element-binding protein, metalloproteinase inhibitor 1 precursor, rap1 GTPase activating protein 1 (RAP1GAP). In conclusion, ER and ENR MDS patients erythroid cells present a different pattern of gene expression with respect to normal. These results may provide the basis for early testing of the genes differentially expressed in ER and ENR patients in order to elucidate their role in the prognostication of rHuEPO response in MDS patients with low endogenous EPO levels.

Sign in / Sign up

Export Citation Format

Share Document