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1.
Talanta ; 236: 122879, 2022 Jan 01.
Artigo em Inglês | MEDLINE | ID: mdl-34635259

RESUMO

We present a sensitive label-free surface enhanced Raman spectroscopy (SERS) method for the discrimination between the recombinant and endogenous human Erythropoietin (EPO) isoforms. The proposed methodology comprises a lectin-functionalised extractor chip for the extraction of the recombinant human EPO (rhuEPO) and the endogenous EPO (enEPO) from blood plasma. The disulfide bond molecular structure of the purified isoforms was modified to chemisorb the biomolecules onto a SERS substrate in a unified orientation, thus maximizing the reproducibility and sensitivity of the SERS measurements. The acquired SERS spectra of the EPO isoforms showed diagnostic Raman bands that allowed for the discrimination between rhuEPO and enEPO. The method was also used for the SERS quantification of rhuEPO and enEPO down to 0.1 pM and 0.5 pM, respectively. The SERS determination of the protein isoforms was cross validated against ELISA. The new SERS method has strong potential for the rapid screening of rhuEPO doping in athletes and for the therapeutic drug monitoring of rhuEPO treatment in cancer patients.


Assuntos
Eritropoetina , Análise Espectral Raman , Humanos , Isoformas de Proteínas , Proteínas Recombinantes , Reprodutibilidade dos Testes
2.
Zhongguo Yi Xue Ke Xue Yuan Xue Bao ; 43(5): 736-742, 2021 Oct.
Artigo em Chinês | MEDLINE | ID: mdl-34728034

RESUMO

Objective To compare the efficacy and safety of cyclosporin A(CsA)and CsA combined with recombined human erythropoietin(rhEPO)in the treatment of patients with chronic aplastic anemia(CAA).Methods Data of 79 patients with CAA treated at Department of Hematology,PUMC Hospital between January 2016 and June 2018 were collected for retrospective analysis.Forty-five patients were treated with CsA+rhEPO,and the other 34 patients with CsA alone.All the enrolled patients were treated for at least 1.5-2.0 years and followed for at least 1.0 year.The efficacy,side effects,long-term outcomes were compared between the two groups,and factors that may influence the efficacy were analyzed.Results The patients treated with CsA+rhEPO included 14 males and 31 females,with a median age of 43(19,73)years old.The median treatment duration of CsA and rhEPO was 26(12,38)and 4(3,6)months,respectively,and the median followed-up time was 24(12,42)months.The patients treated with CsA alone included 16 males and 18 females,with a median age of 36(16,85)years old.The median CsA treatment duration was 24(12,40)months and the median follow-up time was 25(12,40)months.There was no statistical difference in baseline characteristics between the two groups(all P>0.05).After 6 months of treatment,CsA+rhEPO group had higher overall response(OR)rate(55.6% vs. 31.3%;χ 2=4.456,P=0.040)and partial response(PR)rate(53.3% vs. 25.0%;χ2=6.181,P=0.019)than CsA group,and the complete response(CR)rate showed no statistical difference between the two groups(2.2% vs. 6.3%;χ2=0.810,P=0.567).The CR,PR,and OR rates showed no statistical difference between the two groups after 3 and 12 months of treatment and at the end of follow-up(all P>0.05).Similarly,the hemoglobin level at the sixth month of treatment with CsA+rhEPO was higher than that with CsA alone [(102.6±24.0)g/L vs.(90.3±29.1)g/L;t=2.017,P=0.047].However,it showed no significant difference between the two groups at other time points(all P>0.05).The side effects,including an increase in serum creatinine,slight increase in bilirubin,increase in aminotransferase,mild to moderate gingival hyperplasia,gastrointestinal reaction,and muscular fibrillation,had no significant differences between the two groups(all P>0.05),except that 11.1%(5/45)patients in the CsA+rhEPO group reported soreness at the injection site.The two groups showed similar rates of clonal revolution during the follow-up period and no death.No clinical factor was found for prediction of the efficacy of CsA+rhEPO. Conclusion CsA+rhEPO has higher OR rate and hemoglobin level than CsA alone at the sixth month of treatment,and it has similar side effects compared with CsA alone.


Assuntos
Anemia Aplástica , Eritropoetina , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Anemia Aplástica/tratamento farmacológico , Ciclosporina/uso terapêutico , Eritropoetina/uso terapêutico , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Proteínas Recombinantes , Indução de Remissão , Estudos Retrospectivos , Adulto Jovem
3.
J Coll Physicians Surg Pak ; 31(12): 1417-1421, 2021 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-34794280

RESUMO

OBJECTIVE: To compare the effectiveness of erythropoietin alpha and erythropoietin beta in anemia management in the hemodialysis population. STUDY DESIGN: Quasi-experimental study. PLACE AND DURATION OF STUDY: Department of Nephrology, The Kidney Center Postgraduate Training Institute (TKC-PGTI), Karachi, from December 2019 to July 2020. METHODOLOGY: All participants were initially started on erythropoietin alpha and then converted to erythropoietin beta after three months. The effectiveness of the erythropoietin alpha and erythropoietin beta was calculated on the basis of net change of mean hemoglobin and mean hematocrit level in the last four weeks on either erythropoietin therapy. RESULTS: A total of 80 patients completed the study, in which 47 (58.8%) were males while 33 (41.3%) were females. The mean age was 59.7 ± 14.7 years. The net mean hemoglobin change during last 04 weeks was  Ì¶ 0.19 ± 1.2 and  Ì¶ 0.03 ± 1.0 for erythropoietin alpha and erythropoietin erythropoietin beta, respectively (p = 0.41). The net mean hematocrit change during the last four weeks was  Ì¶ 0.45 ± 3.9 and  ̶ 0.49 ± 3.7 for erythropoietin alpha and erythropoietin beta, respectively (p = 0.95). The mean weekly erythropoietin dosage per Kg body weight during the last four weeks was 177.6 ± 130.4 IU/Kg/week for erythropoietin alpha and 121.3 ± 69.6 IU/Kg/week for erythropoietin beta (p = <0.001). CONCLUSION: Erythropoietin alpha and erythropoietin beta have similar therapeutic efficacy in anemia management in chronic kidney disease patients. Reduced dosage of erythropoietin beta achieves and maintains the target hemoglobin level. Key Words: Efficacy, Erythropoietin, Anemia, Chronic kidney disease.


Assuntos
Anemia , Eritropoetina , Falência Renal Crônica , Insuficiência Renal Crônica , Idoso , Anemia/tratamento farmacológico , Anemia/etiologia , Feminino , Hemoglobinas/análise , Humanos , Masculino , Pessoa de Meia-Idade , Proteínas Recombinantes , Diálise Renal , Insuficiência Renal Crônica/complicações , Insuficiência Renal Crônica/tratamento farmacológico
4.
J Biomed Sci ; 28(1): 73, 2021 Nov 02.
Artigo em Inglês | MEDLINE | ID: mdl-34724959

RESUMO

BACKGROUND: Renal erythropoietin (EPO)-producing (REP) cells produce EPO through hypoxia-inducible factor (HIF) 2α-activated gene transcription. Insufficient EPO production leads to anemia in patients with chronic kidney disease. Although recombinant EPO is effective to improve anemia, no reliable REP cell lines limit further progress of research and development of novel treatment. METHODS: We screened Epo mRNA expression in mouse fibroblast cell lines. Small interfering RNA specific for HIF1α or HIF2α was transfected to study Epo expression in C3H10T1/2 cells. The effect of transforming growth factor-ß1 (TGF-ß1) on HIF-EPO axis was studied in C3H10T1/2 cells and mice. RESULTS: Similar to mouse REP pericytes, C3H10T1/2 cells differentiated to α-smooth muscle actin+ myofibroblasts after exposure to TGF-ß1. Specific HIF knockdown demonstrated the role of HIF2α in hypoxia-induced Epo expression. Sustained TGF-ß1 exposure increased neither DNA methyltransferase nor methylation of Epas1 and Epo genes. However, TGF-ß1 repressed HIF2α-encoding Epas1 promptly through activating activin receptor-like kinase-5 (ALK5), thereby decreasing Epo induction by hypoxia and prolyl hydroxylase domain inhibitor roxadustat. In mice with pro-fibrotic injury induced by ureteral obstruction, upregulation of Tgfb1 was accompanied with downregulation of Epas1 and Epo in injured kidneys and myofibroblasts, which were reversed by ALK5 inhibitor SB431542. CONCLUSION: C3H10T1/2 cells possessed the property of HIF2α-dependent Epo expression in REP pericytes. TGF-ß1 induced not only the transition to myofibroblasts but also a repressive effect on Epas1-Epo axis in C3H10T1/2 cells. The repressive effect of TGF-ß1 on Epas1-Epo axis was confirmed in REP pericytes in vivo. Inhibition of TGF-ß1-ALK5 signaling might provide a novel treatment to rescue EPO expression in REP pericytes of injured kidney.


Assuntos
Fatores de Transcrição Hélice-Alça-Hélice Básicos/genética , Eritropoetina/genética , Fator de Crescimento Transformador beta1/genética , Células 3T3 , Animais , Fatores de Transcrição Hélice-Alça-Hélice Básicos/metabolismo , Eritropoetina/metabolismo , Fibroblastos/metabolismo , Camundongos , Camundongos Endogâmicos C57BL , Fator de Crescimento Transformador beta1/metabolismo
5.
J Assoc Physicians India ; 69(10): 11-12, 2021 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-34781656

RESUMO

OBJECTIVES: This study was carried out to evaluate the effect of pegylated erythropoietin and to compare its effects with the effects of darbepoetin alfa on anemia of chronic kidney patients on maintenance hemodialysis having erythropoietin hyporesponsiveness. METHODS: Forty adult patients of chronic kidney disease(CKD) with erythropoietin hyporesponsiveness undergoing maintenance hemodialysis were included in the study. These patients were randomly divided into two groups, Group A consisting of 20 patients who received Subcutaneous Pegylated erythropoietin at a dose of 0.6 mcg/kg body weight, once in every two weeks along with intravenous iron 100 mg/week for 3 months. Group B patients received subcutaneous darbepoietin alfa at a dose of 0.45 mcg/kg body weight once weekly along with iv iron 100mg /week for 3 months. Hematological, renal and inflammatory parameters such as erythrocyte sedimentation rate, C reactive protein, serum ferritin and transferrin saturation were measured at monthly intervals for three months, compiled and analyzed statistically. RESULTS: At the end of the study, in group A there was a significant rise in the hemoglobin, haematocrit and transferrin saturation (p < 0.001 for each of them) while there was a significant decrease in serum ferritin levels (p<0.001). In group &B the increase in hemoglobin, haematocrit and transferrin saturation were not statistically significant (p>0.05), and also there was a significant rise in the serum ferritin levels at the end of the study (p< 0.05). The mean rise in hemoglobin between subsequent months was higher in group A as compared to group B which was statistically significant. CONCLUSION: Pegylated erythropoietin is better than darbepoetin alfa in overcoming erythropoietin hyporesponsiveness and maintaining stable hemoglobin levels in CKD patients on maintenance hemodialysis.


Assuntos
Anemia , Eritropoetina , Falência Renal Crônica , Insuficiência Renal Crônica , Adulto , Anemia/tratamento farmacológico , Anemia/etiologia , Darbepoetina alfa , Hemoglobinas , Humanos , Polietilenoglicóis , Proteínas Recombinantes , Diálise Renal , Insuficiência Renal Crônica/complicações , Insuficiência Renal Crônica/terapia
6.
Anal Chim Acta ; 1185: 339084, 2021 Nov 15.
Artigo em Inglês | MEDLINE | ID: mdl-34711323

RESUMO

Recombinant human erythropoietin (EPO) is a complex therapeutic glycoprotein with three N- and one O-glycosylation sites. Glycosylation of EPO influences its safety and efficacy and is defined as a critical quality attribute. Thus, analytical methods for profiling EPO glycosylation are highly demanded. Owing to the complexity of the intact protein, information about EPO glycosylation is commonly derived from released glycan and glycopeptide analysis using mass spectrometry (MS). Alternatively, comprehensive insights into the glycoform heterogeneity of intact EPO are obtained using ESI MS-based methods with or without upfront separation of EPO glycoforms. MALDI MS, typically performed with TOF mass analyzers, has been also used for the analysis of intact EPO but, due to the poor glycoform resolution, has only provided limited glycoform information. Here, we present a MALDI FT-ICR MS method for the glycosylation profiling of intact EPO with improved glycoform resolution and without loss of sialic acid residues commonly observed in MALDI analysis. Three EPO variants were characterized in-depth and up to 199 glycoform compositions were assigned from the evaluation of doubly-charged ions, without any deconvolution of the mass spectra. Key glycosylation features such as sialylation, acetylation, and N-acetyllactosamine repeats were determined and found to agree with previously reported data obtained from orthogonal analyses. The developed method allowed for a fast and straightforward data acquisition and evaluation and can be potentially used for the high-throughput comparison of EPO samples throughout its manufacturing process.


Assuntos
Eritropoetina , Glicosilação , Humanos , Proteínas Recombinantes , Espectrometria de Massas por Ionização por Electrospray , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz
7.
Medicine (Baltimore) ; 100(42): e27601, 2021 Oct 22.
Artigo em Inglês | MEDLINE | ID: mdl-34678911

RESUMO

BACKGROUND: This study investigated whether administering erythropoiesis-stimulating agents (ESAs) improves endothelial function in patients with non-dialysis chronic kidney disease (CKD) and anemia. METHODS: This single-center, prospective, single-arm comparison study enrolled patients with non-dialysis CKD (stages 4-5) and hemoglobin levels <10 g/dL. ESA administration followed the Kidney Disease: Improving Global Outcomes guideline. The primary endpoint was the change in flow-mediated dilatation after ESA administration in individual patients. The secondary endpoints were changes in 6-minute walk test results, blood pressure, New York Heart Association class, and echocardiographic parameters. The echocardiographic parameters examined included chamber quantification, Doppler parameters, and systolic and diastolic function parameters. RESULTS: Initially, 13 patients were screened, but 2 discontinued due to either heart failure or voluntary withdrawal. The mean flow-mediated dilatation values significantly increased by 10.59% (from 1.36% ±â€Š1.91% to 11.95% ±â€Š8.11%, P = .001). Echocardiographic findings showed that the left ventricular mass index decreased by 11.9 g/m2 (from 105.8 ±â€Š16.3 to 93.9 ±â€Š19.5 g/m2, P  =  .006), and the left atrial volume index decreased by 10.8 mL/m2 (from 50.1 ±â€Š11.3 to 39.3 ±â€Š11.3 mL/m2, P = .004) after 12 weeks of ESA administration. There were no significant differences between pre- and post-ESA treatment 6-minute walk test results. No significant side effects were observed during the study period. CONCLUSIONS: This is the first clinical study to demonstrate that an ESA improves endothelial dysfunction, left ventricular hypertrophy, and left atrial volume in patients with non-dialysis CKD. Thus, ESAs may be considered as adjunctive therapy for reducing cardiovascular risk in these patients.


Assuntos
Anemia/tratamento farmacológico , Anemia/epidemiologia , Endotélio Vascular/efeitos dos fármacos , Eritropoetina/uso terapêutico , Hematínicos/uso terapêutico , Polietilenoglicóis/uso terapêutico , Insuficiência Renal Crônica/epidemiologia , Idoso , Pressão Sanguínea , Comorbidade , Ecocardiografia , Eritropoetina/farmacologia , Feminino , Taxa de Filtração Glomerular , Hematínicos/farmacologia , Hemoglobinas , Humanos , Hipertrofia Ventricular Esquerda , Masculino , Pessoa de Meia-Idade , Polietilenoglicóis/farmacologia , Estudos Prospectivos , Índice de Gravidade de Doença , Teste de Caminhada
8.
Biomater Sci ; 9(19): 6474-6485, 2021 Sep 28.
Artigo em Inglês | MEDLINE | ID: mdl-34582522

RESUMO

Stimulus-responsive polymer materials have attracted much attention as drug carriers because of the ability to deliver drugs to the active site. Reactive oxygen species (ROS) play crucial roles in cellular signaling and regulation of oxygen homeostasis. However, ROS are present in abnormally high levels in many pathological environments. Based on the above points, three-arm poly(lactic-co-glycolic acid)-PO-poly(ethylene glycol) (3s-PLGA-PO-PEG or simply PP) was synthesized by using peroxalate esters (PO) as hydrogen peroxide-responsive linkages. PP was used to deliver promote hematopoietic recovery drugs erythropoietin (EPO) and EPO nanoparticles (EPO NPs) were prepared. We established a hematopoietic system injury model by ionizing radiation (IR) and unexpectedly found the good therapeutic effect of blank PP. Moreover, the administration of EPO NPs obviously decreased IR-induced ROS in bone marrow cells (BMCs) and reconstituted hematopoietic stem cells in BMCs. This study reveals a novel ROS-responsive polymer material that could be employed to remove excess ROS in the lesion and promote the efficacy of drug therapy.


Assuntos
Eritropoetina , Nanopartículas , Portadores de Fármacos , Radiação Ionizante , Espécies Reativas de Oxigênio
9.
Mol Med ; 27(1): 120, 2021 09 26.
Artigo em Inglês | MEDLINE | ID: mdl-34565332

RESUMO

BACKGROUND: Since fall 2019, SARS-CoV-2 spread world-wide, causing a major pandemic with estimated ~ 220 million subjects affected as of September 2021. Severe COVID-19 is associated with multiple organ failure, particularly of lung and kidney, but also grave neuropsychiatric manifestations. Overall mortality reaches > 2%. Vaccine development has thrived in thus far unreached dimensions and will be one prerequisite to terminate the pandemic. Despite intensive research, however, few treatment options for modifying COVID-19 course/outcome have emerged since the pandemic outbreak. Additionally, the substantial threat of serious downstream sequelae, called 'long COVID' and 'neuroCOVID', becomes increasingly evident. Among candidates that were suggested but did not yet receive appropriate funding for clinical trials is recombinant human erythropoietin. Based on accumulating experimental and clinical evidence, erythropoietin is expected to (1) improve respiration/organ function, (2) counteract overshooting inflammation, (3) act sustainably neuroprotective/neuroregenerative. Recent counterintuitive findings of decreased serum erythropoietin levels in severe COVID-19 not only support a relative deficiency of erythropoietin in this condition, which can be therapeutically addressed, but also made us coin the term 'hypoxia paradox'. As we review here, this paradox is likely due to uncoupling of physiological hypoxia signaling circuits, mediated by detrimental gene products of SARS-CoV-2 or unfavorable host responses, including microRNAs or dysfunctional mitochondria. Substitution of erythropoietin might overcome this 'hypoxia paradox' caused by deranged signaling and improve survival/functional status of COVID-19 patients and their long-term outcome. As supporting hints, embedded in this review, we present 4 male patients with severe COVID-19 and unfavorable prognosis, including predicted high lethality, who all profoundly improved upon treatment which included erythropoietin analogues. SHORT CONCLUSION: Substitution of EPO may-among other beneficial EPO effects in severe COVID-19-circumvent downstream consequences of the 'hypoxia paradox'. A double-blind, placebo-controlled, randomized clinical trial for proof-of-concept is warranted.


Assuntos
COVID-19/complicações , COVID-19/tratamento farmacológico , Eritropoetina/genética , Hipóxia/tratamento farmacológico , Pulmão/efeitos dos fármacos , COVID-19/genética , COVID-19/patologia , COVID-19/virologia , Eritropoetina/análogos & derivados , Eritropoetina/uso terapêutico , Humanos , Hipóxia/genética , Hipóxia/patologia , Hipóxia/virologia , Pulmão/patologia , Pulmão/virologia , Pandemias , Proteínas Recombinantes/genética , Proteínas Recombinantes/uso terapêutico , SARS-CoV-2/efeitos dos fármacos
10.
Adv Ther ; 38(10): 5345-5360, 2021 10.
Artigo em Inglês | MEDLINE | ID: mdl-34523074

RESUMO

INTRODUCTION: This integrated phase 3 analysis examined efficacy and cardiovascular safety for roxadustat vs erythropoiesis-stimulating agents (ESAs) in dialysis-dependent patients. METHODS: Efficacy and safety results from four phase 3, randomized, open-label studies comparing roxadustat to ESAs (PYRENEES, SIERRAS, HIMALAYAS, ROCKIES) in dialysis-dependent patients with anemia of chronic kidney disease (CKD) were evaluated by study, pooled population and in two subgroups: incident dialysis and stable dialysis. The primary efficacy endpoint per study was hemoglobin change from baseline (CFB) to weeks 28-36 using least-squares mean difference (LSMD) without rescue therapy. Pooled safety endpoints included time to major adverse cardiovascular event (MACE; myocardial infarction, stroke, and all-cause mortality [ACM]) and MACE+ (MACE plus congestive heart failure or unstable angina requiring hospitalization), ACM, and treatment-emergent adverse events (TEAEs). MACE and MACE+ were evaluated for non-inferiority at 1.8 and 1.3 margins using hazard ratios (HRs) and 95% confidence intervals (CIs). TEAEs were descriptively summarized. RESULTS: In total, 4714 patients were randomized (2354 roxadustat; 2360 ESA). Hemoglobin CFB to weeks 28-36 achieved non-inferiority for roxadustat vs ESA in each study. Roxadustat was non-inferior to ESA for risks for MACE and MACE+ in the entire cohort (MACE: HR 1.09, 95% CI 0.95-1.26; MACE+ : HR 0.98, 95% CI 0.86-1.11) and similar to the incident dialysis and stable dialysis subgroups; ACM results were consistent with MACE and MACE+ (HR 1.13, 95% CI 0.95-1.34). TEAEs were generally comparable between groups. CONCLUSION: Roxadustat improved hemoglobin similarly to ESA while demonstrating comparable cardiovascular and overall safety profiles in a wide spectrum of dialysis-dependent patients with anemia of CKD. Roxadustat represents an oral alternative to ESAs for achieving a target hemoglobin for anemia of CKD in dialysis-dependent patients.


Assuntos
Eritropoetina , Hematínicos , Insuficiência Renal Crônica , Glicina/efeitos adversos , Glicina/análogos & derivados , Hemoglobinas , Humanos , Isoquinolinas/efeitos adversos , Diálise Renal , Insuficiência Renal Crônica/complicações , Insuficiência Renal Crônica/terapia
11.
Nutrients ; 13(9)2021 Sep 18.
Artigo em Inglês | MEDLINE | ID: mdl-34579127

RESUMO

We sought to investigate the effects of resistance training (RT) combined with erythropoietin (EPO) and iron sulfate on the hemoglobin, hepcidin, ferritin, iron status, and inflammatory profile in older individuals with end-stage renal disease (ESRD). ESRD patients (n: 157; age: 66.8 ± 3.6; body mass: 73 ± 15; body mass index: 27 ± 3), were assigned to control (CTL; n: 76) and exercise groups (RT; n: 81). The CTL group was divided according to the iron treatment received: without iron treatment (CTL-none; n = 19), treated only with iron sulfate or EPO (CTL-EPO or IRON; n = 19), and treated with both iron sulfate and EPO (CTL-EPO + IRON; n = 76). The RT group followed the same pattern: (RT-none; n = 20), (RT-EPO or IRON; n = 18), and (RT-EPO + IRON; n = 86). RT consisted of 24 weeks/3 days per week at moderate intensity of full-body resistance exercises prior to the hemodialysis section. The RT group, regardless of the iron treatment, improved iron metabolism in older individuals with ESRD. These results provide some clues on the effects of RT and its combination with EPO and iron sulfate in this population, highlighting RT as an important coadjutant in ESRD-iron deficiency.


Assuntos
Eritropoetina/uso terapêutico , Falência Renal Crônica/terapia , Treinamento de Força , Idoso , Ferritinas/sangue , Compostos Ferrosos/uso terapêutico , Hemoglobinas/análise , Hepcidinas/sangue , Humanos , Inflamação/terapia , Ferro/sangue , Pessoa de Meia-Idade
12.
Int J Mol Sci ; 22(18)2021 Sep 11.
Artigo em Inglês | MEDLINE | ID: mdl-34576001

RESUMO

Perinatal hypoxic-ischemic (HI) brain injury, often in conjunction with an inflammatory insult, is the most common cause of death or disability in neonates. Therapeutic hypothermia (TH) is the standard of care for HI encephalopathy in term and near-term infants. However, TH may not always be available or efficacious, creating a need for novel or adjunctive neurotherapeutics. Using a near-term model of inflammation-sensitized HI brain injury in postnatal day (P) 17 ferrets, animals were randomized to either the control group (n = 43) or the HI-exposed groups: saline vehicle (Veh; n = 42), Ur (uridine monophosphate, n = 23), Epo (erythropoietin, n = 26), or TH (n = 24) to test their respective therapeutic effects. Motor development was assessed from P21 to P42 followed by analysis of cortical anatomy, ex vivo MRI, and neuropathology. HI animals took longer to complete the motor assessments compared to controls, which was exacerbated in the Ur group. Injury resulted in thinned white matter tracts and narrowed cortical sulci and gyri, which was mitigated in Epo-treated animals in addition to normalization of cortical neuropathology scores to control levels. TH and Epo treatment also resulted in region-specific improvements in diffusion parameters on ex vivo MRI; however, TH was not robustly neuroprotective in any behavioral or neuropathological outcome measures. Overall, Ur and TH did not provide meaningful neuroprotection after inflammation-sensitized HI brain injury in the ferret, and Ur appeared to worsen outcomes. By comparison, Epo appears to provide significant, though not complete, neuroprotection in this model.


Assuntos
Eritropoetina/farmacologia , Hipotermia Induzida , Hipóxia-Isquemia Encefálica/terapia , Neuroproteção , Fármacos Neuroprotetores/farmacologia , Uridina/farmacologia , Animais , Modelos Animais de Doenças , Furões , Hipóxia-Isquemia Encefálica/metabolismo , Hipóxia-Isquemia Encefálica/patologia
13.
Int J Mol Sci ; 22(17)2021 Aug 31.
Artigo em Inglês | MEDLINE | ID: mdl-34502385

RESUMO

Erythropoietin (Epo) is the critical hormone for erythropoiesis. In adults, Epo is mainly produced by a subset of interstitial fibroblasts in the kidney, with minor amounts being produced in the liver and the brain. In this study, we used the immortalized renal interstitial fibroblast cell line FAIK F3-5 to investigate the ability of the bioactive sphingolipid sphingosine 1-phosphate (S1P) to stimulate Epo production and to reveal the mechanism involved. Stimulation of cells with exogenous S1P under normoxic conditions (21% O2) led to a dose-dependent increase in Epo mRNA and protein levels and subsequent release of Epo into the medium. S1P also enhanced the stabilization of HIF-2α, a key transcription factor for Epo expression. S1P-stimulated Epo mRNA and protein expression was abolished by HIF-2α mRNA knockdown or by the HIF-2 inhibitor compound 2. Furthermore, the approved S1P receptor modulator FTY720, and its active form FTY720-phosphate, both exerted a similar effect on Epo expression as S1P. The effect of S1P on Epo was antagonized by the selective S1P1 and S1P3 antagonists NIBR-0213 and TY-52156, but not by the S1P2 antagonist JTE-013. Moreover, inhibitors of the classical MAPK/ERK, the p38-MAPK, and inhibitors of protein kinase (PK) C and D all blocked the effect of S1P on Epo expression. Finally, the S1P and FTY720 effects were recapitulated in the Epo-producing human neuroblastoma cell line Kelly, suggesting that S1P receptor-dependent Epo synthesis is of general relevance and not species-specific. In summary, these data suggest that, in renal interstitial fibroblasts, which are the primary source of plasma Epo, S1P1 and 3 receptor activation upregulates Epo under normoxic conditions. This may have a therapeutic impact on disease situations such as chronic kidney disease, where Epo production is impaired, causing anemia, but it may also have therapeutic value as Epo can mediate additional tissue-protective effects in various organs.


Assuntos
Eritropoetina/metabolismo , Receptores de Esfingosina-1-Fosfato/metabolismo , Animais , Fatores de Transcrição Hélice-Alça-Hélice Básicos/metabolismo , Fatores de Transcrição Hélice-Alça-Hélice Básicos/fisiologia , Linhagem Celular , Células Cultivadas , Eritropoese , Eritropoetina/fisiologia , Fibroblastos/metabolismo , Cloridrato de Fingolimode/metabolismo , Humanos , Hipóxia/metabolismo , Hipóxia/fisiopatologia , Rim/metabolismo , Lisofosfolipídeos/metabolismo , Camundongos , Ligação Proteica , Receptores de Lisoesfingolipídeo/metabolismo , Insuficiência Renal Crônica/metabolismo , Transdução de Sinais , Esfingosina/análogos & derivados , Esfingosina/metabolismo , Receptores de Esfingosina-1-Fosfato/fisiologia
14.
Artigo em Inglês | MEDLINE | ID: mdl-34574408

RESUMO

Altitude training increases haemoglobin, erythropoietin values among athletes, but may have negative physiological consequences. An alternative, although less explored, that has the potential to positively influence performance while avoiding some of the negative physiological consequences of hypoxia is sand training. Ten endurance-trained athletes (age: 20.8 ± 1.4, body mass: 57.7 ± 8.2 kg, stature: 176 ± 6 cm; 5000 m 14:55.00 ± 0:30 min) performed three 21-day training camps at different locations: at a high altitude (HIGH), at the sea-level (CTRL), at the sea-level on the sand (SAND). Differences in erythropoietin (EPO) and haemoglobin (Hb) concentration, body weight, VO2max and maximal aerobic velocity (VMA) before and after each training cycle were compared. Data analysis has indicated that training during HIGH elicited a greater increase in VO2max (2.4 ± 0.2%; p = 0.005 and 1.0 ± 0.2%; p < 0.001) and VMA (2.4 ± 0.2%, p < 0.001 and 1.2 ± 0.2%; p = 0.001) compared with CTRL and SAND. While increases in VO2max and VMA following SAND were greater (1.3 ± 0.1%; p < 0.001 and 1.2 ± 0.1%; p < 0.001) than those observed after CTRL. Moreover, EPO increased to a greater extent following HIGH (25.3 ± 2.7%) compared with SAND (11.7 ± 1.6%, p = 0.008) and CTRL (0.1 ± 0.3%, p < 0.001) with a greater increase (p < 0.01) following SAND compared with CTRL. Furthermore, HIGH and SAND elicited a greater increase (4.9 ± 0.9%; p = 0.001 and 3.3 ± 1.1%; p = 0.035) in Hb compared with CTRL. There was no difference in Hb changes observed between HIGH and SAND (p = 1.0). Finally, athletes lost 2.1 ± 0.4% (p = 0.001) more weight following HIGH vs. CTRL, while there were no differences in weight changes between HIGH vs. SAND (p = 0.742) and SAND vs. CTRL (p = 0.719). High-altitude training and sea-level training on sand resulted in significant improvements in EPO, Hb, VMA, and VO2max that exceeded changes in such parameters following traditional sea-level training. While high-altitude training elicited greater relative increases in EPO, VMA, and VO2max, sand training resulted in comparable increases in Hb and may prevent hypoxia-induced weight loss.


Assuntos
Eritropoetina , Corrida , Adulto , Altitude , Hemoglobinas/metabolismo , Humanos , Hipóxia , Consumo de Oxigênio , Adulto Jovem
15.
J Appl Physiol (1985) ; 131(4): 1340-1347, 2021 10 01.
Artigo em Inglês | MEDLINE | ID: mdl-34498946

RESUMO

The aim was to investigate if acute recombinant human erythropoietin (rHuEPO) injection had an effect on mitochondrial function and if exercise would have an additive effect. Furthermore, to investigate if in vitro incubation with rHuEPO had an effect on muscle mitochondrial respiratory capacity. Eight healthy young men were recruited for this double-blinded randomized placebo-controlled crossover study. rHuEPO (400 IU/kg body wt) or saline injection was given intravenously, before an acute bout of exercise. Resting metabolic rate and fat oxidation were measured. Biopsies were obtained at baseline, 120 min after injection, and right after the acute exercise bout. Mitochondrial function (mitochondrial respiration and H2O2 emission) was measured in permeabilized skeletal muscle using high-resolution respirometry and fluorometry. Specific gene expression and enzyme activity were measured. Skeletal muscle mitochondrial respiratory capacity was measured with and without incubation with rHuEPO. Fat oxidation at rest increased after rHuEPO injection, but no difference was found in fat oxidation during exercise. Mitochondrial respiratory capacity was increased after rHuEPO injection when pyruvate was in the assay, which was not the case when saline was injected. No changes were seen in H2O2 emission after rHuEPO injection or acute exercise. Incubation of skeletal muscle fibers in vitro with rHuEPO increased mitochondrial respiratory capacity. Acute rHuEPO injection increased mitochondrial respiratory capacity when pyruvate was used in the assay. No statistical difference was found in H2O2 emission capacity, although a numerical increase was seen after rHuEPO injection. In vitro incubation of the skeletal muscle sample with rHuEPO increases mitochondrial respiratory capacity.NEW & NOTEWORTHY The effect of an acute rHuEPO injection on skeletal muscle mitochondrial function was investigated in young healthy male subjects. rHuEPO has an acute effect on skeletal muscle mitochondrial respiratory capacity in humans, where an increased mitochondrial respiratory capacity was seen. This could be the first step leading to increased mitochondrial biogenesis.


Assuntos
Eritropoetina , Peróxido de Hidrogênio , Estudos Cross-Over , Eritropoetina/metabolismo , Humanos , Peróxido de Hidrogênio/metabolismo , Masculino , Mitocôndrias , Músculo Esquelético/metabolismo
16.
Rinsho Ketsueki ; 62(8): 938-943, 2021.
Artigo em Japonês | MEDLINE | ID: mdl-34497234

RESUMO

Oxygen biology is currently a focus of intensive scientific research. Three scientists received the Nobel prize in physiology or medicine for their outstanding scientific efforts in revealing the mechanisms of oxygen sensing and defense against hypoxia. Hypoxia is a final common pathway to end-stage kidney disease and plays a crucial role in the pathogenesis of cardiovascular complications. Hypoxia-inducible factors (HIFs) are master regulators of defensive mechanisms against hypoxia. Erythropoietin is one of the main targets of HIFs that enhances oxygen delivery by increasing the production of red blood cells. HIF levels are regulated by HIF-prolyl hydroxylase (HIF-PH) inhibitors, which are now available as a new therapeutic modality against anemia in chronic kidney disease. HIF-PH inhibitors raise some theoretical concerns, but should be noted for their potential organ-protective effects.


Assuntos
Anemia , Eritropoetina , Inibidores de Prolil-Hidrolase , Insuficiência Renal Crônica , Anemia/tratamento farmacológico , Anemia/etiologia , Humanos , Hipóxia , Inibidores de Prolil-Hidrolase/uso terapêutico , Insuficiência Renal Crônica/complicações , Insuficiência Renal Crônica/tratamento farmacológico
17.
Nat Commun ; 12(1): 4988, 2021 08 17.
Artigo em Inglês | MEDLINE | ID: mdl-34404781

RESUMO

Glycans are fundamental cellular building blocks, involved in many organismal functions. Advances in glycomics are elucidating the essential roles of glycans. Still, it remains challenging to properly analyze large glycomics datasets, since the abundance of each glycan is dependent on many other glycans that share many intermediate biosynthetic steps. Furthermore, the overlap of measured glycans can be low across samples. We address these challenges with GlyCompare, a glycomic data analysis approach that accounts for shared biosynthetic steps for all measured glycans to correct for sparsity and non-independence in glycomics, which enables direct comparison of different glycoprofiles and increases statistical power. Using GlyCompare, we study diverse N-glycan profiles from glycoengineered erythropoietin. We obtain biologically meaningful clustering of mutant cell glycoprofiles and identify knockout-specific effects of fucosyltransferase mutants on tetra-antennary structures. We further analyze human milk oligosaccharide profiles and find mother's fucosyltransferase-dependent secretor-status indirectly impact the sialylation. Finally, we apply our method on mucin-type O-glycans, gangliosides, and site-specific compositional glycosylation data to reveal tissues and disease-specific glycan presentations. Our substructure-oriented approach will enable researchers to take full advantage of the growing power and size of glycomics data.


Assuntos
Vias Biossintéticas , Glicômica , Polissacarídeos/biossíntese , Transporte Biológico , Vias Biossintéticas/genética , Análise por Conglomerados , Análise de Dados , Eritropoetina/metabolismo , Fucosiltransferases/genética , Gangliosídeos , Técnicas de Inativação de Genes , Glicosilação , Humanos , Mucinas
18.
Drugs Today (Barc) ; 57(8): 491-497, 2021 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-34405206

RESUMO

Anemia is a common complication in patients with chronic kidney disease (CKD). Erythropoiesis-stimulating agents (ESAs) are the standard therapy for anemia in CKD. It has been expected that hypoxia-inducible factor prolyl hydroxylase (HIF-PH) inhibition may have the potential to provide therapeutic benefits over pre-existing ESAs for anemia in CKD. Enarodustat (JTZ-951) is an oral HIF-PH inhibitor. In preclinical studies, enarodustat has been found to increase HIF-alpha proteins, erythropoietin production and erythropoiesis. Enarodustat also shows efficient iron utilization in iron-related parameters during erythropoiesis. Clinical trials have shown that enarodustat improved anemia both in non-dialysis-dependent CKD patients and dialysis patients. The safety results in clinical trials demonstrate that enarodustat is generally well tolerated. On the basis of these results, enarodustat was approved in September 2020 in Japan for the treatment of anemia associated with CKD. This manuscript will review enarodustat, its pharmacological characteristics in preclinical studies, and its efficacy and safety in clinical trials with anemic patients in CKD.


Assuntos
Anemia , Eritropoetina , Insuficiência Renal Crônica , Anemia/tratamento farmacológico , Anemia/etiologia , Humanos , Prolina Dioxigenases do Fator Induzível por Hipóxia , Glicinas N-Substituídas , Piridinas , Insuficiência Renal Crônica/complicações , Triazóis
19.
Commun Biol ; 4(1): 938, 2021 08 05.
Artigo em Inglês | MEDLINE | ID: mdl-34354241

RESUMO

Erythropoietin (EPO) improves neuronal mitochondrial function and cognition in adults after brain injury and in those afflicted by psychiatric disorders. However, the influence of EPO on mitochondria and cognition during development remains unexplored. We previously observed that EPO stimulates hippocampal-specific neuronal maturation and synaptogenesis early in postnatal development in mice. Here we show that EPO promotes mitochondrial respiration in developing postnatal hippocampus by increasing mitochondrial content and enhancing cellular respiratory potential. Ultrastructurally, mitochondria profiles and total vesicle content were greater in presynaptic axon terminals, suggesting that EPO enhances oxidative metabolism and synaptic transmission capabilities. Behavioural tests of hippocampus-dependent memory at early adulthood, showed that EPO improves spatial and short-term memory. Collectively, we identify a role for EPO in the murine postnatal hippocampus by promoting mitochondrial function throughout early postnatal development, which corresponds to enhanced cognition by early adulthood.


Assuntos
Cognição/efeitos dos fármacos , Eritropoetina/administração & dosagem , Hipocampo/efeitos dos fármacos , Mitocôndrias/efeitos dos fármacos , Neurônios/efeitos dos fármacos , Animais , Hipocampo/fisiologia , Camundongos , Camundongos Transgênicos , Mitocôndrias/metabolismo , Neurônios/metabolismo , Distribuição Aleatória
20.
Biomolecules ; 11(8)2021 07 30.
Artigo em Inglês | MEDLINE | ID: mdl-34439791

RESUMO

CHO is the cell line of choice for the manufacturing of many complex biotherapeutics. The constant upgrading of cell productivity is needed to meet the growing demand for these life-saving drugs. Manipulation of small non-coding RNAs-miRNAs-is a good alternative to a single gene knockdown approach due to their post-transcriptional regulation of entire cellular pathways without posing translational burden to the production cell. In this study, we performed a high-throughput screening of 2042-human miRNAs and identified several candidates able to increase cell-specific and overall production of Erythropoietin and Etanercept in CHO cells. Some of these human miRNAs have not been found in Chinese hamster cells and yet were still effective in them. We identified miR-574-3p as being able, when overexpressed in CHO cells, to improve overall productivity of Erythropoietin and Etanercept titers from 1.3 to up to 2-fold. In addition, we validated several targets of miR-574-3p and identified p300 as a main target of miR-574-3p in CHO cells. Furthermore, we demonstrated that stable CHO cell overexpressing miRNAs from endogenous CHO pri-miRNA sequences outperform the cells with human pri-miRNA sequences. Our findings highlight the importance of flanking genomic sequences, and their secondary structure features, on pri-miRNA processing offering a novel, cost-effective and fast strategy as a valuable tool for efficient miRNAs engineering in CHO cells.


Assuntos
Eritropoetina/genética , Etanercepte/metabolismo , Engenharia Genética/métodos , MicroRNAs/genética , Transgenes , Animais , Células CHO , Cricetulus , Proteína p300 Associada a E1A/genética , Proteína p300 Associada a E1A/metabolismo , Eritropoetina/biossíntese , Etanercepte/química , Regulação da Expressão Gênica , Ensaios de Triagem em Larga Escala , Humanos , MicroRNAs/metabolismo , Conformação de Ácido Nucleico , Proteínas Recombinantes/biossíntese , Proteínas Recombinantes/genética , Proteína Supressora de Tumor p53/genética , Proteína Supressora de Tumor p53/metabolismo
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