Effect Of Recombinant Human Erythropoietin Research Articles

Dose-dependent ultrastructural and morphometric alterations after erythropoietin treatment in rat femoral artery vasospasm model

Cerebral vasospasm is the common cause of poor outcome after aneurysmal subarachnoid hemorrhage (aSAH). Although many agents are experimentally and clinicaly used to protect or recover from vasospasm, an effective neurotherapeutic drug is still missing. Erythropoietin (EPO) is recently a promising candidate. The aim of this study is to investigate the dose-dependent effects of recombinant human EPO (rhEPO) on arterial wall in a rat femoral artery vasospasm model. Thirty two animals were divided into four groups: vasospasm without any treatment (group A), vasospasm +250 IU/kg rhEPO group (group B), vasospasm +500 IU/kg rhEPO group (group C), and control group (group D). Rat femoral artery vasospasm model was used. For groups B and C, 7 days of 250 IU/kg and 500 IU/kg intraperitoneal rhEPO in 0.3 ml saline were administered respectively; and for groups A and D, 0.3 ml saline were administered intraperitoneally without any treatment. After 7 days, histological and morphometric analyses were carried out. Vasospasm alone group demonstrated the highest vessel wall thicknesses, comparing to other groups (p < 0.001). While for groups B and C, vessel wall thickness values were significantly higher than the control group (p < 0.001), between these two groups, there was no significant difference achieved (p > 0.05). In our study, there was no significant difference between the two rhEPO treatment groups, but rhEPO treatment was shown to be histologically and morphometrically effective in vasospasm. However, if dosage of EPO treatment is augmented, successful results may be achieved.

Recombinant human erythropoietin counteracts cisplatin-induced visceral hyperalgesia

Cisplatin exerts its cytotoxic effect through distinct DNA lesions, leading to peripheral neuropathy. The risk of sensory neuropathy is a common problem during cancer treatment with cisplatin, leading to somatic hyperalgesia. Yet, data focussing on cisplatin-induced impairment of the autonomic nervous system are limited. The present study was aimed to investigate the effect of recombinant human erythropoietin (rhEPO) on cisplatin-induced visceral hyperalgesia. C57BL/6 mice were treated either with cisplatin (2 mg/kg, once per week) or with cisplatin (2 mg/kg, once per week) plus rhEPO (40 microg/kg, 3 times per week) for 8 weeks. Controls were treated with saline. To quantify the visceromotor response (VMR) at week 9, standardized electrodes were implanted into the external oblique musculature for electromyographic recordings. After that, animals were decapitated and dorsal root ganglia (DRG) was removed for transmission electron microscopy studies. Cisplatin-treated mice showed a significant increase of VMR compared to the controls [(7080 +/- 969) vs (2864 +/- 279); P< 0.001], while rhEPO dramatically counteracted this effect [(2962 +/- 336) vs (7080 +/- 969); P< 0.001)]. Transmission electron microscopy revealed cisplatin-induced structural lesions of nuclear membrane in DRG cells, which could be ameliorated by rhEPO. Erythropoietin can significantly ameliorate the cisplatin-induced visceral hyperplasia and DRG nuclear membrane structure damage in mice, indicating a neuroprotective role of erythropoietin.

Erythropoietin in Combination of Tissue Plasminogen Activator Exacerbates Brain Hemorrhage When Treatment Is Initiated 6 Hours After Stroke

Erythropoietin (EPO), a hematopoietic cytokine, exerts neuroprotective effects in experimental stroke. In the present study, we investigated the effect of recombinant human EPO (rhEPO) in combination with tissue plasminogen activator (tPA) on embolic stroke. Rats subjected to embolic middle cerebral artery occlusion (MCAO) were treated with rhEPO (5000 U/kg) in combination with tPA (10 mg/kg) at 2 or 6 hours after MCAO. Control groups consisted of ischemic rats treated with rhEPO (5000 U/kg) alone, tPA (10 mg/kg) alone, or saline at 2 or 6 hours after MCAO. The combination therapy of rhEPO and tPA initiated 6 hours after MCAO did not reduce the ischemic lesion volume and significantly (P<0.05) increased the incidence of brain hemorrhage measured by frequency of gross hemorrhage and a quantitative spectrophotometric hemoglobin assay compared with rats treated with rhEPO alone and tPA alone. However, when the combination therapy was initiated 2 hours after MCAO, the treatment significantly (P<0.05) reduced the lesion volume and did not substantially increase the incidence of hemorrhagic transformation compared with saline-treated rats. Immunostaining analysis revealed that the combination therapy of rhEPO and tPA at 6 hours significantly (P<0.05) increased matrix metalloproteinase-9, NF-kappaB, and interleukin-1 receptor-associated kinase-1 immunoreactive cerebral vessels compared with rats treated with rhEPO alone and saline. EPO exacerbates tPA-induced brain hemorrhage without reduction of ischemic brain damage when administered 6 hours after stroke in a rat model of embolic MCAO and that matrix metalloproteinase-9, NF-kappaB, and interleukin-1 receptor-associated kinase-1 upregulated by the delayed combination therapy may contribute to augmentation of brain hemorrhage.

Review: Friedreich Ataxia and Erythropoietin

In vitro and in vivo studies have provided evidence for neuroprotective properties of Erythropoietin in neurodegenerative disorders. Although the magnitude of effect is still controversial, very recent findings point to neuronal protection in the central nervous system by Erythropoietins. Erythropoietin is a powerful growth factor which enhances cellular size and ultimatively increases the number of mitochondria. Friedreich Ataxia (FA), an inherited neurodegenerative disorder is caused by a loss of function mutation in the first intron on chromosome 9. FA patients therefore suffer a marked reduction of Frataxin, a mitochondrial protein which is involved in mitochondrial iron homeostasis and/or assembly of iron-sulfur (FeS) proteins and heme synthesis. Mitochondrial dysfunction results in a deleterious energy deficit especially in tissues highly dependent on oxidative phosphorylation such as neurons, muscle cells or pancreatic insular cells. Beneficial effects of recombinant human Erythropoietin (rhuEPO) may derive from an increase in Frataxin levels through currently unknown post-transcriptional and/or post-translational mechanisms. Moreover, additional effects via BDNF and through mitochondrial iron chelation may complete the spectrum of rhuEPOs actions in FA and may be part of its beneficial treatment effects. However, there are clear limitations to chronic rhuEPO treatment. Apart from hematopoietic side effects, tumor growth may be enhanced by rhuEPO application. In this review we provide an overview of studies using rhuEPO in FA and discuss potential beneficial effects of Erythropoietin in FA.

Preventing Negative Effects of Smoking on Microarterial Anastomosis

Although microsurgery has rapid expanded, problems related to microarterial anastomosis continue. Cigarette smoking is one of the major risks for anastomosis by increasing platelet adhesion, and its effects on endothelial cells. Aim of this article is to study the negative effects of cigarettes on microarterial anastomosis line, and to investigate the protective effects of recombinant human erythropoietin (rHuEPO).Ninety-six Sprague-Dawley male rats were divided into 3 groups: group 1 was the control. Rats in groups 2 and 3 were exposed to cigarette smoke starting 21 days prior to surgery for 3 times a day. In group 3, additional 150 IU/kg rHuEPO was given via subcutaneously every 48 hours after microvascular anastomosis, femoral arterial samples, and blood samples were taken for assessment at 1st, 3rd, 5th, and 7th day. Intimae/media ratios were calculated for morphologic analyses.On morphologic analysis of femoral arteries there were statistically significant differences for all 3 groups at 1st, 3rd, 5th, and 7th days (P < 0, 05). The group that made differences was group 2, according to one-way analysis of variance within 3 groups in all days.Smoking decreases endothelial cells healing and causes more thromboses. rHuEPO can prevent these negative effects of smoking.

In vitro and in vivo Cytogenetic Effects of Recombinant Human Erythropoietin on the Frequency of Sister Chromatid Exchanges Alone or in Combination with Mitomycin C

Background: Erythropoietin (EPO) is a glycoprotein which has a main property, erythropoiesis, but its range of action in the human body is very wide. It has been suggested that EPO acts cytoprotectively for many cell lines against many toxic causes in vitro and in vivo. Our aim was to study the action of EPO on DNA of two cell types, human lymphocytes in vitro and on P388 ascites tumor cells inoculated in BDF1 mice in the presence and absence of the genotoxic agent mitomycin C (MMC). Method: The sister chromatid exchange (SCE) assay was used as it is a very sensitive, simple and rapid method for detecting DNA damage. Proliferation rate indices (PRI) and mitotic indices (MI) were also counted. Results: EPO did not alter the SCE level when it acted alone on both cell lines. MMC as a potent genotoxic agent increased SCE levels in vitro and in vivo. EPO used in combination with MMC significantly decreased SCE levels and increased PRI and MI values induced by MMC alone both in vitro and in vivo. Conclusions: EPO acts protectively against the genotoxic potential of MMC, and this action may have clinical implications.

Effect of Recombinant Human Erythropoietin On the Stemness of Bone Marrow-derived Mesenchymal Stem Cells in vitro.

The purpose of this study was to investigate the effects of the recombinant human erythropoietin (rhEPO) on proliferative and multi-differentiation potential of the bone marrow-derived mesenchymal stem cells (MSCs). The MSCs were isolated primarily from bone marrow of adult rat and purified at increasing passage. A purified population of MSCs can be obtained about 2 weeks after the initiation of culture. After three passages (P3-MSCs), bone marrow-derived adherent cells were identified, then different concentrations of rhEPO (0.1, 1, 5, 10, 100 U/ml) was added into the Passage-3 cells which had been identified. The expression of the surface markers in adherent cells was detected by the flow cytometry. The mRNA levels of transcription factors OCT4, SOX2, Nanog and TERT were measured by reverse transcription-polymerase chain reaction (RT-PCR). The results showed that CD29 and CD90 were positive in MSCs, but not CD33, CD44 and CD45, and the cells could differentiate into multiple lineages such as osteocytes and adipocytes. The expression of OCT4, SOX2, TERT, Nanog mRNA were up-regulated by the treatment of EPO. The effect of EPO was the most obvious when its concentration was 5U/mL after 12h. we conclude that MSCs can not only perserve characteristics of stem cells but also maintain its multi-lineage differentiation potential after appropriate treatment of EPO.

Recombinant Human Erythropoietin in the Prevention of Late Anemia in Intrauterine Transfused Neonates With Rh-isoimmunization

The majority of neonates with Rh-isoimmunization develops late anemia between the second and the sixth week of life. We report the effectiveness of recombinant human erythropoietin (rHuEPO) in preventing late anemia in 25 intrauterine and nonintrauterine-transfused neonates. The neonates were treated from 11+/-4 days after birth to 26+/-14 days (400 U/kg/d of rHuEpo, administered subcutaneously). During rHuEpo therapy, vitamin E, calcium folinate, and iron maltose were administered intramuscularly on a daily basis. Hematocrit, platelet, and neutrophil counts did not differ significantly before and after 21-days therapy. However, average values for reticulocyte showed a significant increase. The hematocrit values in the non-intrauterine transfusion (IUT) group increased progressively from the beginning to the end of the treatment, whereas that in the IUT group remained stable. Reticulocyte count increased during treatment in both groups, but it was significantly elevated in the non-IUT group only. Moreover, we observed that only neonates transfused with IUTs needed transfusions before and after treatment. This study suggests the effectiveness of rHuEpo therapy in the treatment of neonates with Rh-isoimmunization and it highlights how IUTs decrease the neonatal response efficacy. Larger, better if multicentric, randomized controlled trial are needed to definitely state whether rHuEPO safely decreases the incidence of late onset anemia.

Effect of Recombinant Human Erythropoietin in a Rat Model of Moderate Chronic Renal Failure - Focus on Inflammation, Oxidative Stress and Function/Renoprotection

Background/Aims: Chronic renal failure (CRF) patients develop anaemia, thus promoting cardiovascular complications, which seems to be favoured by the low kidney erythropoietin (EPO) production. The renal insufficiency degree might determine the moment to start recombinant human EPO (rhEPO) therapy. It has been attributed important non-hematopoietic effects to rhEPO, which might underlie cardio and renoprotection. This work aimed to evaluate the effect of rhEPO in a rat model of moderate CRF, focusing on inflammation, oxidative stress and function/renoprotection. Methods: Four groups (n=7) of male Wistar rats were evaluated during a 15 week follow-up period: control (without treatment); rhEPO (50 IU/Kg/wk Recormon ® ); CRF and CRF+rhEPO. Blood samples were collected at the beginning and 3, 9 and 12 weeks afternephrectomy, in order to evaluate: renal function, haematological parameters, iron metabolism and serum proliferative (TGF- 1), inflammatory (TNF- , CRP, IL-2 and IL-1 ) and redox status (MDA, TAS and 3-NT) markers. Kidney gene expression of Il2, Vegf, Nos2 and Nos3 were assessed by real-time PCR. Blood pressure, heart rate and tissues trophy indexes were also estimated. Results: Our data are consistent with a sustained moderate degree of CRF with development of moderate and corrected anaemia and hypertension. The remnant kidney showed a proliferative profile, with increased mass (hypertrophism), upregulated tissue Vegf gene expression, accompanied by increased levels of serum TGF- 1. Serum 3-NT was augmented, suggesting oxidative stress, which was accompanied by a trend to higher kidney Nos gene expression of both isoforms. rhEPO treatment was able to partially attenuate renal function markers, totally correct anaemia, also demonstrating a proliferative and antioxidant action, suggesting renoprotection. Conclusion: This study suggests that rhEPO therapy might be recommended in moderate CRF stages in order to efficiently correct not only the anaemia but also the underlying deleterious mechanisms, due to a proliferative and antioxidant action on the remnant kidney.

Effect of Recombinant Human Erythropoietin on Serum S100B Protein and Interleukin-6 Levels After Traumatic Brain Injury in the Rat

Erythropoietin (EPO) has a neuroprotective effect in the animal model of ischemia/hypoxia, but the mechanisms underlying the EPO effect in traumatic brain injury (TBI) are not well understood. This study examined the potential neuroprotective mechanisms of recombinant human EPO (rhEPO) in rats after TBI. Sixty healthy adult male Sprague-Dawley rats were randomly divided into 5 groups: 1000 U/kg rhEPO-treated, 3000 U/kg rhEPO-treated, 5000 U/kg rhEPO-treated, citicoline, and normal saline (control) groups. The TBI model was based on the modified Feeney's free falling model. Serum samples were collected at 6 hours, 24 hours, 3 days, 5 days, and 7 days after trauma. The serum S100B protein and interleukin-6 (IL-6) levels were measured after treatment in each group with double antibody sandwich enzyme-linked immunosorbent assay. Both serum S100B protein and IL-6 levels were significantly lower in 3000 U/kg rhEPO-treated and 5000 U/kg rhEPO-treated groups (p < 0.001). The decrease in serum S100B protein level was correlated with the dosage of rhEPO. Medium doses of rhEPO achieved the optimum decreases in the serum IL-6 level. Therefore, inhibition of the composition and secretion of S100B protein and IL-6 levels by EPO might be one of the mechanisms involved in decreasing inflammatory reaction in the brain, and may be responsible for the neuroprotective effect after TBI.

Effects of recombinant human erythropoietin on glutamate expression in the retina with acute high intraocular pressure in a rabbit model

Objective The neuroprotection provided by recombinant human erythropoietin(rhEPO)on the retina from ischemia-reperfusion injury has been confirmed but its mechanism is not fully understood.The present study aimed to investigate the effect of systemic administration of recombinant human erythropoietin(rhEPO)on the expression of glutamate in the retina after acute high intraocular pressure in vitro.MethodsThe acute high intraocular pressure models were established by the perfusion of physiological saline into anterior chamber of the lateral eye in forty-eight Japanese white rabbits.Other 6 Japanese white rabbits were as normal control group.The experimental rabbits were then equally divided into the model group and EPO group,and hypodermic injection of rhEPO was only performed in the EPO group.Glutamate expression in the retina in both groups was observed by immunohistochemistry on days 1,3,7,and 14 after retinal ischemia-reperfusion.Glutamate expression in another 6 rabbit retina without any treatment was determined as normal by the same method.The use of animal followed the Standard of Association for Research in Vision and Ophthalmology.ResultsNo positive expression of glutamate was observed in normal rabbit retina,but positive expression response of glutamate occurred in the rabbit retina of the model group.The number of positive expression cells in the EPO group was more than that in the model group at each time point(P＜0.01).On day 14 after ischemia-reperfusion,the number of positive expression cells was 3.3±1.1 per high visual field in the retina of the model group but 0.3±0.2 in the retina of the EPO group,showing a significant decrease of positive expression cells in EPO group(P＜0.01).ConclusionSystemic administration of rhEPO can down-regulate the expression of glutamate in the retina with acute high intraocular pressure.This process may be one of the mechanisms that rhEPO protects the retina from ischemia reperfusion injury. Key words: erythropoietin; glutamate; retina; neuroprotection

Effects of recombinant human erythropoietin on platelet activation in acute myocardial infarction: Results of a double-blind, placebo-controlled, randomized trial

Erythropoietin mitigates myocardial damage and improves ventricular performance after experimental ischemic injury. This study assessed safety and efficacy markers relevant to the biological activity of recombinant human erythropoietin (rHuEpo) in patients with acute myocardial infarction (MI). We conducted a prospective, placebo-controlled, randomized, double-blind trial to determine the effects of intravenous rHuEpo (200 U/kg daily for 3 consecutive days) on measures of platelet and endothelial cell activation, soluble Fas ligand, and peripheral blood mononuclear cell (PBMC) expression of angiogenesis signaling proteins in 44 subjects with acute MI treated with aspirin and clopidogrel after successful percutaneous coronary intervention. Recombinant human erythropoietin did not alter bleeding time, platelet function assay closure time, von Willebrand factor levels, soluble P-selectin, or soluble Fas ligand levels when compared with placebo. By contrast, rHuEpo significantly increased expression of erythropoietin receptor, vascular endothelial growth factor receptor Flt-1, and phosphorylated phosphatidylinositol 3-kinase in PBMCs when compared with placebo (all Ps < .05). In acute MI patients treated with aspirin and clopidogrel, short-term administration of rHuEpo did not alter markers of platelet and endothelial cell activation associated with thrombosis, yet did increase expression of angiogenesis signaling proteins in PBMCs when compared with placebo. These data provide preliminary evidence of safety and biologic activity of rHuEpo at this dosing and support continued enrollment in ongoing efficacy trials.

PO19-WE-13 Autosomal dominant limb-girdle muscular dystrophy linked to chromosome 7Q36: description of a phenotype

Background: The haematopoietic effect of recombinant human erythropoietin (rhuEPO) in dialysis patients is well known. Recently rhuEPO has received considerable attention because of the finding that it has also broad neuroprotective and cardioprotective capabilities. Nowadays rhuEPO is tested in clinical trials for various neurological conditions such as stroke, schizophrenia ec. where mostly non-anaemic patients are treated with rhuEPO. In this study we investigated the effects of long-term use of rhuEPO in regard to erythropoesis and iron parameters in a group of non-anaemic patients with Friedreich’s ataxia (FA). Patients and methods: In a clinical study 12 adult patients, suffering from the neurodegenerative diseases Friedreich’s ataxia, received 5.000 units rhuEPO 3 times weekly subcutaneously for 8 weeks. Erythropoietic response and parameters of iron metabolism were recorded during rhuEPO treatment and further followed for 10 weeks post treatment. Haematocrit, haemoglobin, reticulocytes, serum iron, serum ferritin, transferrin, transferrin-saturation were monitored using routine laboratory methods. Results: Treatment with rhuEPO showed a significant change in most iron parameters. Interestingly we found a recovery of all iron parameters within 4 to 10 weeks after treatment. Conclusion: Iron parameters in FRDA-patients were significantly changed during eight weeks treatment with rhuEPO, but recovered to baseline levels after completion of the trial. These results indicate that there possibly appears a mobilisation of body iron through rhuEPO treatment.

PO19-WE-12 Treatment with recombinant human Erythropoietin: focus on iron status and possible mechanism for iron redistribution

Background: The haematopoietic effect of recombinant human erythropoietin (rhuEPO) in dialysis patients is well known. Recently rhuEPO has received considerable attention because of the finding that it has also broad neuroprotective and cardioprotective capabilities. Nowadays rhuEPO is tested in clinical trials for various neurological conditions such as stroke, schizophrenia ec. where mostly non-anaemic patients are treated with rhuEPO. In this study we investigated the effects of long-term use of rhuEPO in regard to erythropoesis and iron parameters in a group of non-anaemic patients with Friedreich’s ataxia (FA). Patients and methods: In a clinical study 12 adult patients, suffering from the neurodegenerative diseases Friedreich’s ataxia, received 5.000 units rhuEPO 3 times weekly subcutaneously for 8 weeks. Erythropoietic response and parameters of iron metabolism were recorded during rhuEPO treatment and further followed for 10 weeks post treatment. Haematocrit, haemoglobin, reticulocytes, serum iron, serum ferritin, transferrin, transferrin-saturation were monitored using routine laboratory methods. Results: Treatment with rhuEPO showed a significant change in most iron parameters. Interestingly we found a recovery of all iron parameters within 4 to 10 weeks after treatment. Conclusion: Iron parameters in FRDA-patients were significantly changed during eight weeks treatment with rhuEPO, but recovered to baseline levels after completion of the trial. These results indicate that there possibly appears a mobilisation of body iron through rhuEPO treatment.

Erythropoietin attenuates white matter damage, proinflammatory cytokine and chemokine induction in developing rat brain after intra‐uterine infection

To investigate the possible ameliorating effect of recombinant human erythropoietin (rhEPO) on white matter damage, pro-inflammatory cytokine and chemokine induction in developing rat brain after intra-uterine Escherichia coli infection. E. coli was inoculated into uterine cervix of the time-pregnant rats and the control was injected with normal saline. Following maternal E. coli inoculation, the pups received a single intraperitoneal injection of rhEPO at a dose of 5000 IU/kg body weight immediately after birth. Immunohistochemical staining and Western blot analysis for 2', 3'-cyclic nucleotide 3'-phosphodiesterase (CNPase), neurofilament (NF) and glial fibrillary acidic protein (GFAP) were performed to assess white matter damage in pup brains at post-natal day 1 (P1), P3 and P7. Pro-inflammatory cytokines and chemokines were detected by real-time quantitative RT-PCR at the mRNA levels to evaluate the inflammatory response in pup brains at P1, P3 and P7. A single dose of rhEPO treatment (5000 IU/kg body weight) attenuated white matter damage in developing rat brain after intra-uterine E. coli infection. The protein levels of CNPase and NF in pup brains at P7 significantly increased after post-natal rhEPO treatment as compared with the intra-uterine E. coli-treated group. Also, post-natal rhEPO injection markedly attenuated the intra-uterine E. coli infection-induced increases in GFAP protein expression and the mRNA levels of pro-inflammatory cytokines and chemokines. Post-natal EPO administration as a single dose may exert a neuroprotective effect on white matter damage by reducing pro-inflammatory cytokine and chemokine induction in developing rat brain after intra-uterine E. coli infection.

Effect of recombinant human erythropoietin on insulin resistance in hemodialysis patients

Insulin resistance is a characteristic feature of uremia. Insulin resistance and concomitant hyperinsulinemia are present irrespective of the type of renal disease. Treatment with recombinant human erythropoietin (rHuEPO) was said to be associated with improvement in insulin sensitivity in uremic patients. The aim of this study was to compare insulin resistance in adult uremic hemodialysis (HD) patients including diabetic patients treated with or without rHuEPO. A total of 59 HD patients were studied, patients were divided into 2 groups of subjects: 30 HD patients on regular rHuEPO treatment (group A), and 29 HD patients not receiving rHuEPO (group B) diabetic patients were not excluded. Full medical history and clinical examination, hematological parameters, lipid profile, serum albumin, parathyroid horomone, Kt/V, fasting glucose, and insulin levels were measured in all subjects. Homeostasis Model Assessment of Insulin Resistance (HOMA-IR) was used to compare insulin resistance. The results of this study showed that the mean insulin level of HD patients treated with rHuEPO (group A) (17.5 +/- 10.6 microU/mL) was significantly lower than patients without rHuEPO (group B) (28.8 +/- 7.7 microU/mL), (P<0.001). Homeostasis Model Assessment of Insulin Resistance levels in group A were significantly lower than in group B (3.8 +/- 2.97, 7.98 +/- 4.9, respectively, P<0.001). Insulin resistance reflected by HOMA-IR levels among diabetic patients in group A was significantly lower than among diabetic patients in group B (3.9 +/- 3.2, 9.4 +/- 7.2, respectively, P<0.001). Also, HOMA-IR levels among nondiabetic patients in group A were significantly lower than among nondiabetic patients in group B (3.7 +/- 2.85, 6.9 +/- 1.43, respectively, P<0.01). We found a statistically significant negative correlation between duration of erythropoietin treatment, fasting blood glucose, insulin levels, and insulin resistance (r=-0.62, -0.71, and -0.57, P<0.001). Patients treated with rHuEPO showed less insulin resistance compared with patients not treated with rHuEPO in diabetic and nondiabetic patients and, duration of erythropoietin treatment is negatively correlated with insulin levels and insulin resistance in HD patients.

The Effect of Recombinant Human Erythropoietin on the Development of Retinopathy of Prematurity

In addition to its hematopoietic effects, erythropoietin causes an increased release of endothelin-1 and the stimulation of angiogenesis and thereby it may have possible role in development of retinopathy of prematurity (ROP). Our objective was to determine if an association exists between recombinant human erythropoietin (rhEPO) treatment and the development of ROP. Our case-control study involved 85 very low birthweight infants with birthweights <1500 g born during 2003 and 2004. All the infants were divided into two groups on the basis of whether they got rhEPO or not. The rhEPO was given at the dose of 200 to 250 units/kg/dose three times a week for 10 doses. Further duration of rhEPO therapy was decided on the basis of the clinical response. Ophthalmological examinations were done at the age of 5 to 6 weeks and were repeated 1 to 4 weeks after the first examination according to the severity of the ROP disease during their in-hospital stay. Of 85 infants, 56 (66%) received rhEPO and 29 (34%) did not. In the rhEPO-treated group, 12 infants (21%) had ROP; in the non-rhEPO group, 11 infants (38%) developed ROP. This difference is not statistically significant (odds ratio = 2.63; P = 0.10). There was no correlation between the use of rhEPO and the stage of ROP (random sample = -0.01; P = 0.89). There was no significant difference in the incidence of plus, prethreshold, or threshold disease and the treatment required for ROP between the rhEPO-treated and the nontreated group. The study showed there is no significant difference in the incidence and severity of ROP between the rhEPO-treated and nontreated group.

The Primary Study of CD90(+)CD34(-)and Sca-1(+) Stem Cells Mobilized by EPO Plus G-CSF in Mice.

To investigate effects of recombinant human erythropoietin (rhEPO) plus recombinant human granulocyte colony stimulating factor (rhG-CSF) on mobilization of CD90(+)CD34(-) and Sca-1(+) stem cells in mice. rhEPO 1000 IU·kg(-1)·d(-1) per mice alone or plus rhG-CSF 250 μg·kg(-1)·d-1(-1)per mice was administered to mice for five days, the peripheral blood was collected at 6 hours, 1, 3,5 and 7 days after the last administration. The number of white blood cells (WBC) and mononuclear cells (MNC) was counted. The level of CD34, CD90 and Sca-1 mRNA were detected by reverse transcription-polymerase chain reaction (RT-PCR).The expressions of CD90(+)CD34(-) in absolute nuclear cells of peripheral blood was detected by flow cytometry. The results indicate that compared to mobilizing by rhG-CSF, rhEPO or plus rhG-CSF did not significantly enhance the number of WBC and MNC, on the fifth and the seventh day after the last administration, both the expression of CD34,CD90 and Sca-1 mRNA and the proportion of CD90(+)CD34(-) cells dramatically raised in rhEPO or in addition to rhG-CSF groups. We conclude that rhEPO or plus rhG-CSF had a strong capacity of mobilization of stem cells. The alteration of expression of CD34, CD90 and Sca-1 mRNA after the administration of rhEPO or combined with rhG-CSF indicated that this agents might potentially alter the peripheral blood graft content.

Effects of recombinant human erythropoietin on neuropathic pain and cerebral expressions of cytokines and nuclear factor-kappa B

The effect of recombinant human erythropoietin (rhEPO) on neuropathic pain remains unclear. This study aimed to determine the effects of preemptive administration of rhEPO on the behavioural changes and neuroinflammatory responses in a rat model of neuropathic pain. Fifty rats were randomly allocated into five groups, sham-operation treated with saline and L5 spinal nerve transection treated with different doses of rhEPO (0 [saline], 1000, 3000, or 5000 U x kg(-1), respectively). The rats were intraperitoneally treated from 1 day before surgery to post-surgery day 7. The mechanical (paw pressure thresholds, PPT) and thermal thresholds (paw withdrawal latencies, PWL) were measured on post-surgery days 1, 3, and 7. The contralateral brain was obtained on post-surgery day 7 to determine the expressions of tumour necrosis factor (TNF-alpha), interleukin (IL)-1beta, IL-6, L-10, and nuclear factor-kappa B (NF-kappaB) activity. There were significant decreases in PPT and PWL after L5 spinal nerve transection (P < 0.001). Compared with the saline group, the rhEPO 3000 and 5000 U x kg(-1) groups resulted in significant increases in PPT and PWL (P < 0.001) and reduced the cerebral expressions of TNF-alpha, IL-1beta, IL-6, and NF-kappaB activity associated with the increase in IL-10 (rhEPO3000 group, P < 0.05, and rhEPO5000 group, P < 0.001, respectively). Administration of rhEPO 1000 U x kg(-1) had no significant effects on these variables. Preemptive rhEPO dose-dependently attenuated the mechanical and thermal hyperalgesia in L5 spinal nerve transection rats, which correlated with the decreased cerebral expressions of TNF-alpha, IL-1beta, and IL-6 via downregulating NF-kappaB activity and the increased expression of IL-10.

Effectiveness of recombinant human erythropoietin, vitamin D3and iron therapy on long-term survival of patients with end-stage renal disease receiving haemodialysis: analysis of 702 patients after 10-year follow-up

Few studies have been conducted to investigate the influence of recombinant human erythropoietin (rhEPO) on the long-term prognosis of end-stage renal disease (ESRD). A retrospective cohort study. The largest regional hospital renowned for haemodialysis in northern Taiwan. A total of 702 ESRD patients undergoing haemodialysis between 1993 and 2002 were evaluated. The rate of overall use of rhEPO, vitamin D3 or Fe therapy was 62 %. The 10-year survival rate in patients with rhEPO supplementation was statistically more favourable than that in patients without rhEPO (hazard ratio (HR) = 0.38, 95 % CI 0.30, 0.47, P < 0.0001). Similar findings were noted for patients receiving vitamin D3 (HR = 0.36, 95 % CI 0.21, 0.64, P = 0.0004) and Fe (HR = 0.45, 95 % CI 0.33, 0.61, P < 0.0001). After adjusting for age, education and aetiology, the administration of rhEPO resulted in statistically significant improvements in long-term survival rate either with (HR = 0.30, 95 % CI 0.22, 0.42) or without (HR = 0.48, 95 % CI 0.38, 0.61) combined use of Fe or vitamin D3. We demonstrated a reduction in long-term mortality related to supplementation therapy with rhEPO, vitamin D3 and Fe. The findings provide a justification for the administration of combined supplement therapy in patients undergoing haemodialysis.