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使用中风体外模型研究神经促红细胞生成素的神经保护作用。

Neuroprotective Effects of neuroEPO Using an In Vitro Model of Stroke.

作者信息

Fernando Garzón, Yamila Rodríguez, Cesar García Julio, Ramón Rama

机构信息

Department of Animal Health, University of Nariño, Pasto 52001, Nariño, Colombia.

Institute of Basic and Preclinical Sciences "Victoria de Girón", Havana 10400, Cuba.

出版信息

Behav Sci (Basel). 2018 Feb 13;8(2):26. doi: 10.3390/bs8020026.

Abstract

Erythropoietin (EPO) is a glycoprotein initially identified as a hormone synthesized and secreted by the kidney that regulates erythropoiesis. EPO, and a group of its derivatives, are being evaluated as possible neuroprotective agents in cerebral ischemia. The objective of this study, using an in vitro model, was to determine how neuroEPO-which is a variant of EPO with a low sialic acid content-protects neurons from the toxic action of glutamate. Primary neuronal cultures were obtained from the forebrains of Wistar rat embryos after 17 days of gestation. Excitotoxicity was induced after nine days of in vitro culture by treatment with a medium containing 100 µM glutamate for 15 min. After this time, a new medium containing 100 ng of neuroEPO/mL was added. Morphological cell change was assessed by phase-contrast microscopy. Oxidative stress was analysed by measuring antioxidant and oxidant activity. After 24 h, the treatment with 100 ng of neuroEPO/mL showed a significant ( < 0.01) decrease in mortality, compared to cells treated with glutamate alone. neuroEPO treatment decreased mortality and tended to reproduce the morphological characteristics of the control. The oxidative stress induced by glutamate is reduced after neuroEPO treatment. These results confirm that neuroEPO has a protective effect against neuronal damage induced by excitotoxicity, improving antioxidant activity in the neuron, and protecting it from oxidative stress.

摘要

促红细胞生成素(EPO)是一种糖蛋白,最初被鉴定为肾脏合成和分泌的一种调节红细胞生成的激素。EPO及其一组衍生物正在作为脑缺血中可能的神经保护剂进行评估。本研究使用体外模型的目的是确定神经EPO(一种唾液酸含量低的EPO变体)如何保护神经元免受谷氨酸的毒性作用。原代神经元培养物取自妊娠17天的Wistar大鼠胚胎的前脑。体外培养9天后,用含有100μM谷氨酸的培养基处理15分钟诱导兴奋毒性。此后,加入含有100 ng神经EPO/mL的新培养基。通过相差显微镜评估细胞形态变化。通过测量抗氧化剂和氧化剂活性分析氧化应激。24小时后,与单独用谷氨酸处理的细胞相比,用100 ng神经EPO/mL处理显示死亡率显著降低(<0.01)。神经EPO处理降低了死亡率,并倾向于重现对照的形态特征。神经EPO处理后,谷氨酸诱导的氧化应激降低。这些结果证实神经EPO对兴奋毒性诱导的神经元损伤具有保护作用,可改善神经元中的抗氧化活性,并保护其免受氧化应激。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/cbe2/5836009/f62ff542ce51/behavsci-08-00026-g001.jpg

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