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红细胞生成素受体研究进展 总被引:2,自引:0,他引:2
红细胞生成素受体是促红细胞生成素的作用受体 ,属于细胞因子超家族的成员。近年来 ,由于其克隆表达技术的应用使人们在对造血机制、EPO的信号转导机制 ,及利用其受体筛选 EPO类似物方面的研究有了新的进展。本文综合近年来有关文献对 EPO受体的研究作一概括性介绍。 相似文献
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Ogle Jane W. Lange R. D. Dunn C. D. R. 《In vitro cellular & developmental biology. Plant》1978,14(11):945-950
Summary The in vitro production of the important regulatory of erythropoiesis, erythropoietin (Epo), is reviewed. It is concluded
that it is possible to produce almost routinely small quantities of Epo in tissue culture. Although such procedures offer
the potential to provide large quantities of the hormone for clinical use, the optimum culture conditions and mechanisms for
triggering Epo production have yet to be resolved.
This work was supported by Grants No. 74444 from The John A. Hartford Foundation, and HL 10567 from the National Institutes
of Health. 相似文献
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The frequency of bone marrow cells that bind erythropoietin 总被引:1,自引:0,他引:1
The frequencies of rat and mouse bone marrow cells capable of binding erythropoietin were studied by both direct fluorescence and indirect immunofluorescence. We found that between 1-2% of the cells bound erythropoietin, that the binding was specific, and that the number of cells that bound erythropoietin was, in part, a function of the erythropoietic state of the donor animal. A statistical method for evaluating the data obtained is included. 相似文献
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A monoclonal antibody produced by hydridoma cell line, ATCC HB8209, was used to detect and purify erythropoietin synthesized in a cell-free system. The antibody was raised against the N-terminal 20 residues of erythropoietin. It retained anti-erythropoietin activity in 6 M urea in which most of the cell-free synthesized erythropoietin became soluble and gave an enhanced activity of the antibody. 相似文献
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The purpose of this study was to investigate whether erythropoietin (EPO) and environmental enrichment (EE) after hypoxic ischemic insult can prevent brain damage, improve neuronal plasticity, and reduce sensorimotor deficits and whether the combination of EPO and EE has a synergistic effect. Sixty Sprague-Dawley rats were used for this study. The rats were divided into three groups: middle cerebral artery occlusion (MCAO) without treatment, MACO?+?EPO, and MCAO?+?EPO?+?EE. For the behavior test, the foot-fault test and measurement of the crossing time in the parallel bars were used. Infarct volume measurement, Neuronal nuclei (NeuN) immunohistochemistry, western blot of the synaptophysin (SYP), and Growth-Associated protein 43 (GAP-43) were used. On postoperative days (POD) at 1 week and 4 weeks, group of EPO and EE increased the recovery as seen in the behavior test. The infarct volume of the EPO group was 32.4%, the EPO?+?EE group was 28.5%, and the control group was 44.0% on POD 4 weeks. The SYP and GAP-43 expression showed an increase in the EPO and EPO?+?EE group compared with the MCAO group. EPO and EPO?+?EE treatment creates a neurorestorative effect, improved neural plasticity, and reduced sensorimotor deficits. But the synergistic effect of the combination of EPO and EE was significantly not proved. In the results of this study, we suggest that EPO and EE treatment may offer a neruorestoration, which can improve the functional brain disturbance of stroke patients. 相似文献
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Mónica Mattio Natalia Ceaglio Marcos Oggero Norma Perotti Ignacio Amadeo Gustavo Orozco Guillermina Forno Ricardo Kratje Marina Etcheverrigaray 《Biotechnology progress》2011,27(4):1018-1028
Although historically used for the treatment of anemia, erythropoietin (EPO) has emerged as a neurotrophic and neuroprotective agent in different conditions of neuronal damage (traumatic brain injury, ischemia, spinal cord compression, peripheral neuropathy, retinal damage, epilepsy, Parkinson's Disease, among others). Nonetheless, EPO's therapeutic application is limited due to its hematological side‐effects. With the aim of obtaining EPO derivatives resembling the hormone isolated from cells and tissues of neural origin, a novel combination of less acidic EPO glycoforms ‐designated as neuroepoetin (rhNEPO)‐ was purified to homogeneity from the supernatant of a CHO‐producing cell line by a four‐step chromatographic procedure. This simple and single process allowed us to prepare two EPO derivatives with distinct therapeutic expectations: the hematopoietic version and a minimally hematopoietic, but mainly in vitro cytoprotective, alternative. Further biological characterization showed that the in vivo erythropoietic activity of rhNEPO was 25‐times lower than that of rhEPO. Interestingly, using different in vitro cytoprotective assays we found that this molecule exerts cytoprotection equivalent to, or better than, that of rhEPO in cells of neural phenotype. Furthermore, despite its shorter plasma half‐life, rhNEPO was rapidly absorbed and promptly detected in the cerebrospinal fluid after intravenous administration in rats (5 min postinjection, in comparison with 30 min for rhEPO). Therefore, our results support the study of neuroepoetin as a potential drug for the treatment of neurological diseases, combining high cytoprotective activity with reduced hematological side‐effects. © 2011 American Institute of Chemical Engineers Biotechnol. Prog., 2011 相似文献
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目的:探讨促红细胞生成素(Epo)对大鼠脑缺血/再灌注损伤的保护作用。方法:32只SD大鼠,采用夹闭双侧颈总动脉30min再灌注24h制作脑缺血/再灌注模型。随机分为4组(n=8):假手术组、脑缺血/再灌注组、Epo组及阳性对照组(尼莫地平),观察缺血/再灌注后血清一氧化氮(NO)和脑组织匀浆中超氧化歧化酶(SOD)活性、丙二醛(MDA)含量及脑组织含水量的变化。结果:Epo组血清NO和脑组织匀浆中MDA含量显著下降,SOD活性显著升高,脑组织含水量显著下降,与缺血/再灌注组相比有显著性差异。结论:大鼠脑缺血/再灌注后,Epo能减轻脑组织的含水量,减少自由基的生成,减轻脂质过氧化反应,对脑缺血/再灌注损伤有保护作用。 相似文献
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Construction of a fusion protein between N-terminal 153 peptide of thrombopoietin and erythropoietin
Thrombopoietin(TPO)isahematopoieticcytokineclonedinrecentyears[1—3].Itsfunctionistoregulatetheformationofplatelet,whichplaysanimportantroleinbloodclotting,bystimulatingthestemcellstodifferentiateanddevelopintomaturemegakaryocytes.Theresultsofanimalexperim… 相似文献
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Production of erythropoietin by BHK cells growing on the microcarriers trapped in alginate gel beads
Baby hamster kidney (BHK) cells engineered to produce recombinant human erythropoietin (EPO) were cultured at high density on microcarriers entrapped by calcium alginate gel particles. In this system, the BHK cells proliferated not only on the microcarriers but also in vacant spaces in the alginate gel particles. These spaces contributed greatly to high-density cultivation of the cells and a high productivity of EPO.Abbreviations BHK
Baby Hamster Kidney
- EPO
Erythropoietin 相似文献
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The widespread use of aluminum (Al) provides easy exposure of humans to the metal and its accumulation remains a potential problem. In vivo and in vitro assays have associated Al overload with anemia. To better understand the mechanisms by which Al affects human erythrocytes, morphological and biochemical changes were analyzed after long-term treatment using an in vitro model. The appearance of erythrocytes with abnormal shapes suggested metal interaction with cell surface, supported by the fact that high amounts of Al attached to cell membrane. Long-term incubation of human erythrocytes with Al induced signs of premature erythrocyte death (eryptosis), such as phosphatidylserine externalization, increased intracellular calcium, and band 3 degradation. Signs of oxidative stress, such as significant increase in reactive oxygen species in parallel with decrease in the amount of reduced glutathione, were also observed. These oxidative effects were completely prevented by the antioxidant N-acetylcysteine. Interestingly, erythrocytes were also protected from the prooxidative action of Al by the presence of erythropoietin (EPO). In conclusion, results provide evidence that chronic Al exposure may lead to biochemical and morphological alterations similar to those shown in eryptosis induced by oxidant compounds in human erythrocytes. The antieryptotic effect of EPO may contribute to enhance the knowledge of its physiological role on erythroid cells. Irrespective of the antioxidant mechanism, this property of EPO, shown in this model of Al exposure, let us suggest potential benefits by EPO treatment of patients with anemia associated to altered redox environment. 相似文献
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Weise A Altmann F Rodriguez-Franco M Sjoberg ER Bäumer W Launhardt H Kietzmann M Gorr G 《Plant biotechnology journal》2007,5(3):389-401
The highly glycosylated peptide hormone erythropoietin (EPO) plays a key role in the regulation of erythrocyte maturation. Currently, marketed EPO is produced by recombinant technology in mammalian cell cultures. The complementary DNA (cDNA) for human EPO (hEPO) was transiently and stably expressed in the moss Physcomitrella patens wild-type and Δ-fuc-t Δ-xyl-t mutant, the latter containing N -glycans lacking the plant-specific, core-bound α1,3-fucose and β1,2-xylose. New expression vectors were designed based on a Physcomitrella ubiquitin gene-derived promoter for the expression of hEPO cDNA. Transient expression in protoplasts was much stronger at 10 than at 20 °C. In Western blot analysis, the molecular size of moss-produced recombinant human EPO (rhEPO) was identified to be 30 kDa, and it accumulated in the medium of transiently transformed protoplasts to high levels around 0.5 µg/mL. Transgenic Physcomitrella Δ-fuc-t Δ-xyl-t mutant lines expressing EPO cDNA showed secretion of rhEPO through the cell wall to the culture medium. In 5- and 10-L photobioreactor cultures, secreted rhEPO accumulated to high levels above 250 µg/g dry weight of moss material after 6 days. Silver staining of rhEPO on sodium dodecylsulphate-polyacrylamide gel electrophoresis (SDS-PAGE) taken from the bioreactor culture demonstrated a high purity of the over-expressed secreted rhEPO, with a very low background of endogenous moss proteins. Peptide mapping of rhEPO produced by the Physcomitrella Δ-fuc-t Δ-xyl-t mutant indicated correct processing of the plant-derived signal peptide. All three N -glycosylation sites of rhEPO were occupied by complex-type N -glycans completely devoid of the plant-specific core sugar residues fucose and xylose. 相似文献
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Amin Ghafuri-Esfahani Rahman Shokri Athar Sharifi Lida Shafiee Roya Khosravi 《Preparative biochemistry & biotechnology》2020,50(8):834-841
AbstractSeveral factors may affect erythropoietin (EPO) sugar structures including designing cell culture procedure, pH, concentration of additives, dissolved oxygen, and other physicochemical parameters. In this study, we investigated the influence of changes in effective parameters and compounds on the growth rate of Chinese hamster ovary cell (CHO) cells producing recombinant EPO. Cell culture was performed at different temperature, buffering conditions, and varied concentrations of additives such as pyruvic acid, insulin, GlutaMAX, and sodium butyrate. Results indicated that the optimal temperature and pH were 37?°C and 7.2, respectively. Also, optimal concentrations for pyruvic acid, butyrate, glutamate, and insulin were obtained to be 20?mM, 1?mM, 2?mM, and 40?μg/mL, respectively. Then, cell culture was performed in microcarrier-coated spinner flasks under the optimized condition. The results showed recombinant human EPO (rhEPO) production with adequate purity. Optimization of physicochemical conditions and culture media are important factors to improve the quantity and quality of protein products. This study showed that cell growth and recombinant EPO protein production significantly increased under the optimized conditions. The results of this research can also be used in scale-up to increase the efficiency of EPO production. Abbreviations: EPO: erythropoietin; CHO cell: Chinese hamster ovary cell; rhEPO: recombinant human EPO; DMEM: modified eagle’s medium; FBS: fetal bovine serum; SDS-PAGE: sodium dodecyl sulfate–polyacrylamide gel electrophoresis; IGF-1: insulin-like growth factor 1 相似文献