Erythropoietin Treatment Group Research Articles

Erythropoietin Attenuates Experimental Contrast-Induced Nephrology: A Role for the Janus Kinase 2/Signal Transducer and Activator of Transcription 3 Signaling Pathway.

The aim of the present study was to investigate the effect of erythropoietin (EPO) on contrast-induced nephrology (CIN) in vivo and in vitro. Male C57BL/6J mice were divided into four groups: control, CIN (iohexol 6.0 g/kg), EPO (3,000 IU/kg), and CIN+EPO. Hematoxylin and eosin (H&E) staining and biochemical index analyses were performed to evaluate renal injury. The cellular proliferation rate was detected using the Cell Counting Kit-8 (CCK-8) assay. In addition, a terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) assay and flow cytometric assay were used to assess the apoptosis of tissue and cells, respectively. Renal protein expression associated with apoptosis, pyroptosis, and signaling pathways was determined by Western blot (WB) assays for tissues and cells. The results showed that EPO significantly decreased serum creatinine, blood urea nitrogen, and cystatin C levels and alleviated renal histological changes in vivo. The protein levels of Janus kinase 2/signal transducer and activator of transcription 3 (JAK2/STAT3) signaling pathway components were overexpressed in the EPO treatment group. Furthermore, EPO suppressed the cell apoptosis and pyroptosis; decreased the protein levels of cleaved caspase-3, Bax, gasdermin D (GSDMD), and caspase-1; and enhanced the expression of Bcl-2. In summary, EPO could exert renoprotective effect by activating the JAK2/STAT3 signaling pathway, which may be a novel potential therapy for the treatment of CIN in the clinic.

Lack of association between erythropoietin treatment and risk of depression in patients with end-stage kidney disease on maintenance dialysis: a nationwide database study in Taiwan.

Background:Patients with end-stage kidney disease (ESKD), have been associated with higher risk of developing depression. Erythropoietin (EPO), frequently used for the treatment of anemia in ESKD patients, has been shown to have neuroprotective and antidepressant effects. In this study, we examined whether EPO treatment changed the risk of depression in ESKD patients.Methods:In a nationwide population-based cohort in Taiwan from 1998 to 2013, patients with a diagnosis of ESKD on maintenance dialysis and aged greater than 18 years were classified into EPO treatment group or non-EPO treatment group. All patients were followed up until the diagnosis of depressive disorder or the end of the study period.Results:In this cohort (13,067 patients in the EPO and 67,258 patients in the non-EPO group), 5569 patients were diagnosed as depressive disorder in the follow-up period. We found the risk of depression in EPO group was not significantly different from that in non-EPO group (adjusted hazard ratio = 0.98, 95% confidence interval 0.92–1.04, p = 0.499) after adjusting for sex, age, certification year of catastrophic illness for ESKD, physical co-morbidities, and use of benzodiazepines.Conclusion:In summary, using the nationwide reimbursement data in Taiwan, we found that EPO treatment in ESKD patients was not associated with their general risk of developing depression.

Potential Efficacy of Erythropoietin on Reducing the Risk of Mortality in Patients with Traumatic Brain Injury: A Systematic Review and Meta-Analysis

Objective The objective of this study is to assess the effectiveness of erythropoietin (EPO) on mortality, neurological outcomes, and adverse event in the treatment of traumatic brain injury (TBI). Methods We searched databases including PubMed, OVID, and the Cochrane Library from inception until October 18, 2019 for randomized controlled trials (RCTs) to compare EPO treatment group and placebo in patients with TBI. Two authors independently processed the data and evaluated the quality of inclusion studies. Statistical analysis was performed with heterogeneity test with I2 and chi-square tests. We summarized the mortality, prognosis of neurological function, and deep vein thrombosis (DVT) outcomes and presented as risk ratio (RR) or risk difference (RD) with a 95% CI. Results Seven RCTs accounting for 1180 patients were included after meeting the inclusion criteria. Compared with placebo, the overall mortality of EPO-treated patients was significantly reduced (RR 0.68 [95% CI 0.50-0.93]; p = 0.02). EPO therapy did not improve neurological prognosis (RR 1.21 [95% CI 0.93-1.15]; p = 0.16) or increase the occurrence of DVT (RR 0.83 [95% CI 0.61–1.13]; p = 0.242), which showed no significant difference. Conclusions The results showed that the administration of EPO may reduce the risk of mortality without enhancing the occurrence of DVT in TBI patients. However, the effect of EPO on neurological outcome remains indistinct. Through subgroup analysis, we demonstrated that the dose of EPO may be a potential factor affecting the heterogeneity in neurological function and that the follow-up duration may influence the stability of the result.

Therapeutic effect of erythropoietin on brain injury in premature mice with intrauterine infection.

ObjectiveThe objective of this study is to explore the protective effect of erythropoietin (EPO) on brain injury induced by intrauterine infection in premature infants and its related mechanism, so as to provide reference for clinical medication.MethodsIntrauterine infection model is established by injecting lipopolysaccharide into pregnant mice, and HE staining of mouse placenta is used to judge whether the model of intrauterine infection is successful or not. Fifteen female rats are successfully pregnant and divided into intrauterine infection group (10 rats) and control group (5 rats). The mice in the intrauterine infection group are intraperitoneally injected with lipopolysaccharide (LPS) at a dose of 0.3 mg/kg. After delivery, 16 newborn mice in the control group are randomly selected as blank control group. 32 newborn mice in the intrauterine infection group are selected as model group, and then divided into infection group and EPO treatment group, 16 mice in each group. After birth, mice in the blank control group are intraperitoneally injected with 0.2 mL saline daily. The infected mice are intraperitoneally injected with 0.2 mL saline daily. The mice in the EPO treatment group are intraperitoneally injected with recombinant human erythropoietin (rhEPO) 5000 IU/kg daily. HE staining results, EPOR protein and NMDAR1 mRNA expression in brain tissue of three groups of neonatal mice were compared.ResultsFirstly, the blood vessels of the mice in the intrauterine infection group are markedly hyperemic and edematous, and the infiltration of neutrophils is increased. The white matter structure of the neonatal mice in the intrauterine infection group is loose and stained lightly. The nerve fibers in the brain are different in thickness and disordered in arrangement. The nucleus is small and dark stained. The number of glial cells in brain tissue increases significantly. Secondly, the EPOR protein expression and physiological level of neonatal mice in intrauterine infection group increase significantly at 3, 7 and 14 days after birth. Compared with the blank control group, the difference is statistically significant (P < 0.05). On the 3rd day after birth, the expression level of EPOR protein in the EPO treated group is significantly higher than that in the intrauterine infection group (P < 0.05). Thirdly, the expression level of NMDA R1mRNA in brain tissue of neonatal mice at birth, on the 3rd and 7th day after EPO treatment is significantly lower than that of intrauterine infection group (P < 0.05).ConclusionEPO can promote the proliferation and differentiation of brain endogenous neural stem cells, and has a certain therapeutic effect on brain injury of premature mice caused by intrauterine infection.

Neutropenia in Myelosuppressed Cancer Patients Treated with Recombinant Human Erythropoietin (EPO)

Recombinant human erythropoietin (EPO) has been used extensively for 3 decades to treat various forms of anemia, including anemia associated with cancer treatment. Multiple randomized, placebo controlled clinical trials conducted since the late 1980's found that EPO treatment ameliorated anemia, reduced transfusion requirements, and improved quality of life measures in cancer patients receiving myelosuppressive chemotherapy. However, results of several of these trials suggested that EPO treatment might also exacerbate neutropenia in these patients. This possible side effect had been anticipated by a pre-clinical observation that neutrophil production in long-term marrow cultures was suppressed by EPO. However, chemotherapy-induced neutropenia was expected in these clinical trials and not studied, nor reported, in detail. Recently, primary data from 5 prospective randomized, placebo controlled clinical trials of EPO treatment in cancer patients undergoing myelosuppressive chemotherapy, sponsored by Johnson &amp; Johnson, became available through the Yale Open Data Access (YODA) project*. These data allowed us to compare serial neutrophil counts (ANCs) from 1650 study subjects with various forms of cancer who participated in these 5 clinical trials and were randomly assigned to either EPO or placebo treatment. Individual clinical trial sample sizes varied from 71 to 922 (Table below). ANC values were recorded weekly in 2 trials but less frequently in the other 3. In our analysis we determined the percent of study subjects with recorded ANC values &lt;1500/mm3, &lt;500/mm3, and &lt;200/mm3 during the initial 12 weeks of each trial for individuals assigned either to placebo or to EPO treatment. Mantel-Haenszel odds ratios were used as measures of differences between treatment groups. In addition, Poisson regression was used to assess differences in the total numbers of ANC measurements &lt;500/mm3 per study subject per 12 weeks on study that had been recorded in each treatment group. In each trial more study subjects assigned to EPO than to placebo treatment were found to have had at least one ANC &lt;500/mm3 reflecting severe neutropenia (Table). Study-specific odds ratios for this difference ranged from 1.24 to 2.08 and the Mantel-Haenszel odds ratio for the combined studies was 1.48 with a 95% Confidence Interval (CI) of 1.08 - 2.03, p =0.014. A statistically significant odds ratio was also found for ANC values of &lt;200/mm3 (1.73, 1.10-2.70, p=0.016), but not for neutropenic ANC values overall, i.e. &lt;1500/mm3 (1.18, 0.93-1.50, p=0.17). Total numbers of recorded ANC values &lt;500/mm3 per study subject per 12 weeks on study were also significantly different - 0.66 (95% CI, 0.57-0.76) in the EPO treatment group vs. 0.49 (0.42-0.60) in the placebo group, p=0.0075. This analysis of 5 randomized placebo controlled clinical trials carried out from 1988 to 2002 finds that EPO treatment of individuals undergoing myelosuppressive chemotherapy was associated with statistically significant increases in the incidence of severe neutropenia and suggests that competition between the erythroid and myeloid lineages may occur in the setting of suppressed hematopoietic reserves when the erythroid lineage is pharmacologically amplified. *This study, carried out under YODA Project 2017-2486, used data obtained from the Yale University Open Data Access Project, which has an agreement with JANSSEN RESEARCH &amp; DEVELOPMENT, L.L.C.. The interpretation and reporting of research using this data are solely the responsibility of the authors and does not necessarily represent the official views of the Yale University Open Data Access Project or JANSSEN RESEARCH &amp; DEVELOPMENT, L.L.C.. Table Disclosures No relevant conflicts of interest to declare.

Erythropoietin protects propofol induced neuronal injury in developing rats by regulating TLR4/NF-κB signaling pathway abstract

ObjectiveTo explore the protective effect of erythropoietin on propofol induced neuronal injury in developing rats by regulating TLR4/NF-κB signaling pathway. MethodRats were divided into normal control group (Control), propofol group (PPF), erythropoietin group (EPO), propofol + erythropoietin group (P + E), propofol + TAK-242 group (P + T), and propofol + EPO + LPS group (P + E+L) (n = 12). The pathology of hippocampal neurons was observed. The inflammatory factors were detected by ELISA. The expression of TLR4/NF-κB pathway-related proteins were detected by Western blot. ResultsCompared with the PPF group, the percentage of apoptotic cells in the hippocampal CA1 area of the erythropoietin treatment groups were greatly decreased while the percentage of positive cells in the hippocampus were remarkably increased(p < 0.05). The production of inflammatory factors and the expressions of TLR4/NF-κB pathway-related proteins were greatly improved in treatment groups (p < 0.05). Compared with P + E group, the percentage of apoptotic cells in CA1 area of the P + E+L group was significantly increased and the percentage of positive cells was remarkably reduced(p < 0.05), the levels of interleukin-1β(IL-1β), interleukin-6(IL-6), interleukin-8(IL-8) and tumor necrosis factor-α(TNF-α) were significantly increased and interleukin-4(IL-4) and interleukin-10(IL-10) was notably reduced(p < 0.05). The protein expression of TLR4 and p-P65 was significantly increased, while the protein expression of p-IκBαwas significantly decreased (p < 0.05). ConclusionErythropoietin has a protective effect on propofol induced neuropathic injury propofol in rats, and its mechanism is relevant to the regulation of TLR4/NF-κB signaling pathway.

The Effect of Systemic Erythropoietin Treatment on Retinal Nerve Fiber Layer Parameters in Patients With Chronic Renal Failure Undergoing Peritoneal Dialysis

To evaluate the effect of erythropoietin (EPO) treatment on retinal nerve fiber layer (RNFL) parameters in patients with chronic renal failure (CRF) undergoing peritoneal dialysis (PD). Fifty-eight eyes of 29 patients with CRF undergoing PD were evaluated. Fifteen patients have been treated with EPO (group 1), 14 patients without EPO treatment (group 2), and 30 eyes of 15 age-matched normal control subjects were assessed in group 3. A complete ophthalmologic examination and RNFL measurements were performed for each patient after PD. Anemia parameters were also measured. RNFL thickness protocol was used to acquire circular scans of 3.4 mm in diameter around optic nerve. RNFL thicknesses were evaluated in 4 quadrants. Only the left eyes were recruited for statistical analysis. The mean and quadrantal RNFL thickness values in group 1 were compared with those of groups 2 and 3. The mean RNFL thickness values in patients undergoing PD were statistically lower than the control group at superior, inferior, nasal, and temporal quadrant, respectively (P=0.03, 0.04, 0.04, and 0.03). Differences between the RNFL thickness values in group 1 and group 2 were statistically significant only in the temporal quadrant (P=0.02). In patients with CRF undergoing PD, RNFL thickness parameters were found to be significantly reduced. The effect of EPO on RNFL parameters was statistically significant only in the temporal quadrant.

Transplantation of erythropoietin gene-transfected umbilical cord mesenchymal stem cells as a treatment for limb ischemia in rats.

The aim of this study was to investigate the mechanisms of erythropoietin (EPO)-transfected umbilical cord mesenchymal stem cells (UC-MSCs) in the treatment of rats with ischemic limb and provide a theoretical basis for optimization of UC-MSC transplantation. Sixty SD rats were randomly divided into four groups: ischemia control group, EPO treatment group, UC-MSCs treatment group, EPO gene transfected UC-MSC treatment groups (15 rats in each group). The left femoral hind artery and its branches were ligated to develop hind limb ischemia in male SD rats. Five points were injected in the adductor and gastrocnemius muscles with medium, cDNA3-EPO gene DNA-liposome complex solution or UC-MSCs in control groups and EPO-transfected-UC-MSCs in the experimental group. Western blot confirmed in vitro EPO expression in EPO gene-transfected human UC-MSCs. Arterial angiography at 4 weeks post-transplantation showed no development of blood vessels in the control group and moderate angiogenesis in the EPO- and UC-MSC-treated groups. However, a large number of freshmen angiogenesis and abundant collateral circulation was observed in the EPO-transfected-UC-MSC-treated experimental group. Rat capillary density measurement results confirmed the angiographs quantitatively and showed no statistically significant difference between EPO- and UC-MSC-treated groups (P > 0.05). CM-Dil-positive cell numbers were (0 ± 0.00), (0 ± 0.00), (32.46 ± 6.68), (59.64 ± 10.38)/HP (P < 0.05). RT-PCR detected that the in vivo mRNA expression of the EPO gene was relatively higher in the EPO-transfected-UC-MSC-treated group than the EPO-treated group (0.79 ± 0.06 vs 0.19 ± 0.04, P < 0.05). Thus, this study revealed that using UC-MSCs as vector in gene therapy for limb ischemia facilitates stable and effective expression of EPO compared to direct gene injection.

Erythropoietin activates the phosporylated cAMP [adenosine 3′5′ cyclic monophosphate] response element-binding protein pathway and attenuates delayed paraplegia after ischemia-reperfusion injury

Paraplegia remains a devastating complication of complex aortic surgery. Erythropoietin (EPO) has been shown to prevent paraplegia after ischemia reperfusion, but the protective mechanism remains poorly described in the spinal cord. We hypothesized that EPO induces the CREB (cAMP [adenosine 3'5' cyclic monophosphate] response element-binding protein) pathway and neurotrophin production in the murine spinal cord, attenuating functional and cellular injury. Adult male mice were subjected to 4 minutes of spinal cord ischemia via an aortic and left subclavian cross-clamp. Experimental groups included EPO treatment 4 hours before incision (n = 7), ischemic control (n = 7), and shams (n = 4). Hind-limb function was assessed using the Basso motor score for 48 hours after reperfusion. Spinal cords were harvested and analyzed for neuronal viability using histology and staining with a fluorescein derivative. Expression of phosphorylated (p)AKT (a serine/threonine-specific kinase), pCREB, B-cell lymphoma 2, and brain-derived neurotrophic factor were determined using immunoblotting. By 36 hours of reperfusion, EPO significantly preserved hind-limb function after ischemia-reperfusion injury (P < .01). Histology demonstrated preserved cytoarchitecture in the EPO treatment group. Cords treated with EPO expressed significant increases in pAKT (P = .021) and pCREB (P = .038). Treatment with EPO induced expression of both of the neurotrophins, B-cell lymphoma 2, and brain-derived neurotrophic factor, beginning at 12 hours. Erythropoietin-mediated induction of the CREB pathway and production of neurotrophins is associated with improved neurologic function and increased neuronal viability following spinal cord ischemia reperfusion. Further elucidation of EPO-derived neuroprotection will allow for expansion of adjunct mechanisms for spinal cord protection in high-risk thoracoabdominal aortic intervention.

Impacts of erythropoietin combined with granulocyte‐colony stimulating factor on neural function after traumatic brain injury in rats (618.6)

Objective: To investigate the effect of erythropoietin (EPO) and granulocyte‐colony stimulating factor (G‐CSF) combination therapy on functional recovery and nerve cell regeneration after TBI. Methods: The rats model with TBI was prepare by Feeney’s methods. 100 male Wistar rats were randomly divided into Sham operation group(Sham group), traumatic brain injury group (TBI group), EPO treatment group(T+E group ), G‐CSF treatment group(T+G group), and EPO combined G‐CSF group(T+E+G group), and then each group was sub‐divided into 5 subgroups according to the time 1,3 ,5,7 and 14 days after TBI.(Each subgroups n=4). Each treatment group rats were immediately given corresponding intervention measures after TBI for 3 days. Neurological function was assessed using a modified neurological severity score(mNSS) before rats were sacrificed at 1,3,5,7 and 14 days post TBI . Immunofluorescence measurement was used to detect newly proliferation cells number and its direction of differentiation by BrdU single positive cells and BrdU/NeuN or BrdU/GFAP double positive cells. Results: The mNSS scores of each treatment group was significantly lower than TBI group, and T+E+G group was also significantly lower than T+E group or T+G group at 5 ,7 and 14 days of surgery (P &lt; 0.05); The BrdU positive cells number in T+E+G group was significantly higher than other groups starting from 3 days after TBI, and the peak at 7 days (P &lt; 0.05). Compare the groups of brain tissue BrdU/NeuN and BrdU/GFAP double positive cells number, T+E+G group were significantly higher than other groups, T+E group and T+G group were significantly higher than Sham group and TBI group (P &lt; 0.05). Conclusions: Combined use of EPO and G‐CSF can obviously improve functional recovery and promote the brain nerve cell regeneration differentiating to neurons and astrocytes in the injury boundary zone.Grant Funding Source: This study was supported by the National Natural Science Foundation Youth (Fund No.30600637)

Effects of amino acids and albumin on erythropoietin carbamoylation

Anemia in chronic renal failure results from inadequate production of erythropoietin and decrease in its biological activity, and the reduced activity of erythropoietin is caused by the presence of plasma inhibitors of erythropoietin. It is reported that one of the inhibitors of erythropoietin is cyanate, a potential uremic toxin formed spontaneously from increased urea due to decreased renal function, and erythropoietin loses its biological activity due to negatively charged cyanate. The purpose of this study is to investigate the protective effects of amino acids, positively charged amino groups, and albumin binding of several toxins on erythropoietin carbamoylation. The degree of change in erythropoietin structure by cyanate was measured by the trinitrobenzenesulphonate reaction and Western blotting. The loss of biological activity of erythropoietin caused by cyanate was measured by injecting erythropoietin into rats with chronic renal failure. The free amino groups in erythropoietin decreased under cyanate treatment in a time- and concentration-dependent manner. In the cyanate treatment group, of the twenty amino acids, phenylalanine, valine, tryptophan, threonine, and lysine prevented the structural modification of erythropoietin, according to Western blot analysis. In addition, of the three proteins, albumin prevented the structural modification of erythropoietin. As for the cyanate with erythropoietin treatment group, only lysine and albumin prevented the loss of biological activity of erythropoietin in the rats. The results of this study suggest that lysine and albumin may play a protective role against renal anemia by erythropoietin carbamoylation in chronic renal failure.

Efficacy of Erythropoietin Combined With Enteral Nutrition for the Treatment of Anemia in Crohn's Disease

Anemia is a common and serious complication in patients with inflammatory bowel disease. The present study was dedicated to evaluate the therapeutic efficacy of erythropoietin (EPO) combined with enteral nutrition (EN) in anemic Crohn's disease (CD) patients, in terms of hemoglobin level, treatment success rate, adverse events, and predictor of this therapy. We performed a prospective study in CD patients. On the basis of hemoglobin level, all enrolled patients were divided into anemic and nonanemic groups. The anemic group was further divided into EPO and non-EPO subgroups, depending on whether EPO was prescribed. Hematological and other parameters were measured initially and in the first 4 weeks after starting treatment. In total, 109 patients (49 nonanemic and 60 anemic, including 38 EPO and 22 non-EPO) were included. The prevalence of anemia in CD was 55.05%. Age, disease behavior, Crohn's Disease Activity Index scores, C-reactive protein, and erythrocyte sedimentation rate were significantly different between anemic and nonanemic groups. An increase in hemoglobin level and a significant decrease in C-reactive protein level were observed in the EPO treatment group. Treatment success rate was 63.16% in the EPO group, whereas none of patients achieved treatment success in the non-EPO group. EPO combined with EN can improve the hemoglobin level in anemic CD patients.

The impact of erythropoietin on inflammation and neuronal apoptosis after sciatic nerve injury in rats

Objective To explore the effect of erythropoietin on inflammation, neuronal apoptosis and the possible mechanism after sciatic nerve injury in rats and provide experimental basis for clinical treatment of peripheral nerve injury.Methods Left sciatic nerve defect was created in 36 female Sprngue-Dawley rats which were divided into three groups:normal saline(NS)group,erythropoietin(EPO)group and nerve growth factor(NGF)group.NS,EPO and NGF were administered by intraperitoneal injection in the NS group,EPO group and NGF group,respectively.At 7 and 14 days after the operation,the morphological change of L5 dorsal root ganglion neurons was observed by light microscope.The expression of IL-6 and TNF-α mRNA of both proximal and distal stumps were quantified using RT-PCR.The neuronal apoptosis of L5 dorsal root ganglion was assessed by TUNEL.Results At 7 and 14 days after the operation,the expression of IL-6 mRNA of both proximal and distal stumps in EPO group was less than that in the NS group(P ＜ 0.01).The expression of IL-6 mRNA of distal stumps in EPO group was less than that in the NGF group(P ＜ 0.01).The expression of TNF-α mRNA of both proximal and distal stumps in EPO group was less than that in the NS and NGF groups(P ＜0.01)7 days postoperatively.The expression of TNF-α mRNA of distal stumps in EPO group was less than that in the NS group(P ＜0.05)14 days postoperatively.At 7 and 14 days after the operation,neuronal apoptosis in EPO treatment group was lower than NS treatment group(P ＜ 0.01),while neuronal apoptosis in EPO treatment group was lower than NGF treatment group(P ＜ 0.05)14 days postoperatively.Conclusion EPO can attenuate injury-induced inflammation and inhibit dorsal root ganglion apoptosis,which may provide protective effect for sciatic nerve injury in rats. Key words: Sciatic nerve; Apoptosis; Inflammation; Erythropoietin; Injuries

Protective effects of erythropoietin in experimental spinal cord injury by reducing the C/EBP-homologous protein expression

Objectives: Erythropoietin (EPO) is a variety of tissue-protective functions, including spinal cord. This study aimed to determine the neuron protective effect of erythropoietin on spinal cord injury (SCI) by assessing C/EBP-homologous protein (CHOP) in the development of a rat model of SCI.Methods: Sixty Sprague–Dawley rats were randomly assigned to three groups: sham-operation control group, SCI group, and EPO treatment group. By using a weight-drop contusion SCI model, the rats in the SCI group and EPO treatment group were killed at 1 and 7 days subsequently. The Basso, Beattie, and Bresnahan (BBB) scores were examined for locomotor function. Pathological changes were observed after hematoxylin–eosin (H&E) staining. The expression of CHOP was determined by immunohistochemical staining and RT-PCR analysis.Results: BBB scores showed more quick recovery in the erythropoietin treatment group than that in the SCI group (P<0·01). Pathological changes also revealed a reduction in the volume of cavitations and more neurons regeneration in the EPO treatment rats than that of the SCI rats. The number of CHOP positive cells in the SCI group on day 1 and 7 days after SCI increased compared with the erythropoietin treatment group and sham-operation control group (P<0·01). CHOP mRNA folds in sham-operation control rat from 1 to 7 days showed the same trend.Conclusions: Endoplasmic reticulum (ER) stress was triggered at the early stage of SCI. Increased expression of CHOP can be found in the injured segment of the spinal cord after injury. EPO treatment could prevent pathological alterations from severe spinal cord injury by reducing expression of CHOP.

Efficacy of recombinant erythropoietin combined with enteral nutrition for the treatment of anemia in Crohnʼs disease: A single-center experience of 109 Cases

This study was designed to evaluate the efficacy of recombinant erythropoietin (EPO) combined with enteral nutrition in anemic Crohn's disease (CD) patients, considering changes of hemoglobin levels, clinical responses rates and predictors of this combination therapy. We performed a retrospective single-center study in CD patients. Based on the hemoglobin level and treatment, all enrolled patients were divided into anemic (EPO and non-EPO groups) and non-anemic groups. Haematological and biological serological parameters were measured before treatment (baseline) and in the initial four weeks after starting treatment. Totally, 109 patients (49 non-anemic, 60 anemic including 38 EPO and 22 non-EPO) were included. The prevalence of anemia in CD was 55.5%, and age, disease behavior, CDAI scores, C-reactive protein and erythrocyte sedimentation rate were significantly different between anemic and non-anemic groups. Correlation between hemoglobin level and CDAI score was demonstrated by linear regression analysis. A significant increase of hemoglobin level was observed in EPO treatment group, whereas a significant decrease was confirmed in non-EPO group. Treatment success rate was 63.16% in EPO group, whereas none of patients reached clinical response in non-EPO group. None of the haematological and biological serological parameters was associated with or predictor of treatment success. EPO combined with enteral nutrition can improve the hemoglobin level in anemic CD patients. None of the assumed factors could be a biomarker of treatment success.

Abstract 8534: Combination of Cyclosporine and Erythropoietin Improves Brain Infarct Size and Neurological Function in Rats after Ischemic Stroke

Objectives: This study tested the superiority of combined cyclosporine (CsA)-erythropoietin (EPO) therapy compared with either one in limiting brain infarction area (BIA) and preserving neurological function in rat after ischemic stroke (IS). Methods: Fifty adult-male SD rats were equally divided into sham control (group 1), IS plus intra-peritoneal physiological saline (at 0.5/24/48h after IS) (group 2), IS plus CsA (20.0 mg/kg at 0.5/24h, intra-peritoneal) (group 3), IS plus EPO (5,000IU/kg at 0.5/24/48h, subcutaneous) (group 4), combined CsA and EPO treatment (group 5) after occlusion of distal left internal carotid artery. Results: BIA on day 21 after acute IS was highest in group 2 and lowest in group 5 (all p&lt;0.01). The sensorimotor functional test showed highest frequency of left turning in group 2 and lowest in group 5 (all p&lt;0.05). mRNA and protein expressions of apoptotic markers and number of apoptotic nuclei on TUNEL were highest in group 2 and lowest in group 1 and 5, whereas the anti-apoptotic markers exhibited an opposite trend (all p&lt;0.05). The expressions of inflammatory and oxidized protein were highest in group 2 and lowest in group 1 and 5, whereas anti-inflammatory markers showed reversed changes in group 1 and 5 (all p&lt;0.05). The number of aquaporin - 4+ and glial fibrillary acid protein+ stained cells were highest in group 2 and lowest in groups 1 and 5 (all p&lt;0.01). Conclusion: Combined treatment with CsA and EPO was superior to either one alone in protecting rat brain from ischemic damage after IS (the proposed mechanisms of potential therapeutic impact of combined therapy with CsA and EPO on acute IS have been summarized in the figure)

Combination of cyclosporine and erythropoietin improves brain infarct size and neurological function in rats after ischemic stroke

BackgroundThis study tested the superiority of combined cyclosporine A (CsA)-erythropoietin (EPO) therapy compared with either one in limiting brain infarction area (BIA) and preserving neurological function in rat after ischemic stroke (IS).MethodsFifty adult-male SD rats were equally divided into sham control (group 1), IS plus intra-peritoneal physiological saline (at 0.5/24/48 h after IS) (group 2), IS plus CsA (20.0 mg/kg at 0.5/24h, intra-peritoneal) (group 3), IS plus EPO (5,000IU/kg at 0.5/24/48h, subcutaneous) (group 4), combined CsA and EPO (same route and dosage as groups 3 and 4) treatment (group 5) after occlusion of distal left internal carotid artery.ResultsBIA on day 21 after acute IS was higher in group 2 than in other groups and lowest in group 5 (all p < 0.01). The sensorimotor functional test showed higher frequency of left turning in group 2 than in other groups and lowest in group 5 (all p < 0.05). mRNA and protein expressions of apoptotic markers and number of apoptotic nuclei on TUNEL were higher in group 2 than in other groups and lowest in group 1 and 5, whereas the anti-apoptotic markers exhibited an opposite trend (all p < 0.05). The expressions of inflammatory and oxidized protein were higher in group 2 than in other groups and lowest in group 1 and 5, whereas anti-inflammatory markers showed reversed changes in group 1 and other groups (all p < 0.05). The number of aquaporin-4+ and glial fibrillary acid protein+ stained cells were higher in group 2 as compared to other groups and lowest in groups 1 and 5 (all p < 0.01).Conclusioncombined treatment with CsA and EPO was superior to either one alone in protecting rat brain from ischemic damage after IS.

Effect of erythropoietin on the mobility of bone marrow mesenchymal stem cells to the sites of injured spinal cord

Objective To obsevre the effect of erythropoietin (EPO) treatment on the mobility of bone marrow mesenchymal stem cell (BMSCs) to the sites of injured spinal cord and explore the molecular mechanism. Methods Thirty SD rats were randomly divided into sham-operation group, saline group and EPO treatment group. BMSCs labeled with Hoechst33342 were transplanted by tail vein. Functional outcome measurements were evaluated by the Basso, Beattie and Bresnahan (BBB) score at 1st, 3rd, 7th,14th, 21st and 28th day. The immigrating number of BMSCs was measured by immunofluorescent staining.The expression of brain-derived neurotrophic factor (BDNF) and nerve growth factor (NGF) in BMSCs was detected by Western blotting. Results From the 3rd day on, the BBB scores in EPO treatment group were (5.29±0.69), (8.18±0.38), (13.32±0.77), (15.25 ±1.83), (18.71 ±1.54) respectively, which were obviously higher than in saline group. The number of BMSCs was (90. 12 ±7. 68), (116. 26 ± 13. 54),( 186. 32 ± 20. 35 ), (211.64 ± 31.83 ) and (298.71 ± 21.54) respectively, which was significantly increased in EPO treatment group (P ＜ 0. 05 ). The protein levels of BDNF and NGF expressed by BMSCs were increased significantly (P ＜ 0. 05) in EPO treatment group as compared with sham-operation group.Conclusion EPO treatment after spinal cord injury may help BMSCs migrate to the injury site and accelerate recovery of neural function by up-regulating the expression of BDNF and NGF. Key words: Erythropoietin; Bone marrow mesenchymal stem cell; Spinal cord injury

Protective effect of erythropoietin on myocardial infarction in rats by inhibition of caspase-12 expression

In the present study, the myocardial protective effects of erythropoietin (EPO) by inhibition of the expression of caspase-12 were investigated in a myocardial infarction rat model. Thirty male SD rats were divided into three groups: sham-operation group, myocardial infarction group and EPO treatment group. The myocardial infarction model was created by ligating the left anterior descending coronary artery. The EPO treatment group was established by injecting rh-EPO (1,000 IU/kg) intraperitoneally every day after the operation, and the other two groups were injected with sodium chloride. Four weeks after induction of myocardial infarction, the left ventricular diastolic pressure (LVDP) was tested by Langendorff apparatus and the pathological changes were analyzed by H&E staining. Caspase-12 expression in the left ventricular myocardium was also measured by immunohistochemistry. Four weeks after induction of myocardial infarction, the improvement in heart function in the EPO treatment group was more distinct compared to that of the myocardial infarction group; LVDP was higher in the EPO treatment group compared to the myocardial infarction group, but lower compared to the control group. H&E staining showed that the myocardial cells in the normal control group were aligned in order with a clear structure and were stained equably, while the myocardial cells in the myocardial infarction model rats lined up in disorder with an augmented cell body appearing to have many granules and interstitial fibrosis. Myocardial fibrosis and disorder were improved in the EPO treatment group. The expression of caspase-12 in the myocardial infarction group was also increased compared to the EPO treatment group rats. The results suggest that EPO improves heart function in myocardial infarction rats by down-regulating the expression of caspase-12, which may protect the myocardium by abrogating endoplasmic reticulum stress-mediated cardiomyocyte apoptosis and improving heart function.

Investigation of the protective effect of erythropoietin on spinal cord injury in rats

Erythropoietin (EPO) is a promising therapeutic agent used in a variety of spinal cord injuries. Therefore, identifying the specific molecular pathway mediating the neuronal protective effect of EPO after spinal cord injury (SCI) is of great value to the patients concerned. Platelet-derived growth factor (PDGF)-B is an important factor in the recovery of neurological function. We explored changes in the expression of PDGF-B in spinal cord injury rats after receiving EPO treatment. We used a weight-drop contusion SCI model, and EPO treatment group rats received single doses of EPO (1,000 U/kg i.p.) immediately after the operation. Seven days after the operation, the results revealed a more rapid recovery as noted by the higher BBB scores, less disruption and more neuronal regeneration of the spinal cord in the EPO treatment group than that in the SCI group. PDGF-B expression also increased in the EPO treatment group compared to that in the SCI group (P<0.01). This study showed that PDGF-B plays a role in the neuronal protective effect of EPO on spinal cord injury in rats, which may help to explain the quick recovery after EPO treatment of spinal cord injury.