Adembri Chiara, Massagrande Alessandra, Tani Alessia, Miranda Marco, Margheri Martina, De Gaudio Raffaele, Pellegrini-Giampietro Domenico E
Section of Anesthesiology and Intensive Care, Department of Critical Care Medicine, University of Florence, Florence, Italy.
Crit Care Med. 2008 Mar;36(3):975-8. doi: 10.1097/CCM.0B013E3181644343.
The well-documented neuroprotective effects of recombinant human erythropoietin (rhEPO) are commonly associated with untoward erythrocyte-stimulating effects (polycythemia), with subsequent risk of thromboembolic complications. A carbamylated-rhEPO (CEPO) derivative, which is neuroprotective but lacks hematopoietic activity, has been recently developed. In this study, we evaluated the neuroprotective capability of CEPO in an in vitro model of cerebral trauma in which rhEPO was previously shown to reduce posttraumatic cell death.
Prospective, controlled experiment.
Animal, basic science laboratory.
Wistar rats, 8 days old.
Organotypic hippocampal slices, obtained from rat brains, were subjected to a well-characterized model of mechanical injury followed by addition of 10 IU/mL rhEPO, 10-100 IU/mL CEPO, or vehicle (injured control) to the incubation medium at different times to assess the temporal window of therapeutic neuroprotection.
Posttraumatic cell death was quantified at 12, 24, or 48 hrs after injury by measuring propidium iodide fluorescence in the selectively vulnerable CA1 hippocampal area. Posttraumatic injury, observed in injured, vehicle-treated hippocampal slices, was significantly attenuated by addition of either 10 IU/mL rhEPO or 10 IU/mL CEPO. The neuroprotective efficacy of 10 IU/mL rhEPO or CEPO remained intact even when administration was delayed 1 hr after trauma. Qualitative microscopy in semithin sections showed that both rhEPO and CEPO exerted a marked pyramidal neuron-sparing effect.
Our study shows that 10 IU/mL CEPO exerts neuroprotective effects comparable with those of rhEPO in an in vitro model of mechanical cerebral trauma. Because CEPO lacks hematopoietic effects and seems to possess a prolonged therapeutic time window, this erythropoietin derivative may represent an exciting new pharmacologic tool in treating patients with mechanical injury to the brain.
重组人促红细胞生成素（rhEPO）具有充分文献记载的神经保护作用，但其通常与不良的促红细胞生成作用（红细胞增多症）相关，继而存在血栓栓塞并发症的风险。最近研发出一种氨甲酰化 - rhEPO（CEPO）衍生物，它具有神经保护作用但缺乏造血活性。在本研究中，我们在脑外伤的体外模型中评估了CEPO的神经保护能力，在此模型中rhEPO先前已被证明可减少创伤后细胞死亡。
前瞻性对照实验。
动物基础科学实验室。
8日龄的Wistar大鼠。
从大鼠脑获取的海马脑片培养物，接受一个特征明确的机械损伤模型，随后在不同时间向孵育培养基中添加10 IU/mL rhEPO、10 - 100 IU/mL CEPO或赋形剂（损伤对照），以评估治疗性神经保护的时间窗。
通过测量选择性易损的海马CA1区碘化丙啶荧光，在损伤后12、24或48小时对创伤后细胞死亡进行定量。在接受赋形剂处理的损伤海马脑片培养物中观察到的创伤后损伤，通过添加10 IU/mL rhEPO或10 IU/mL CEPO可显著减轻。即使在创伤后1小时延迟给药，10 IU/mL rhEPO或CEPO的神经保护功效仍然完好。半薄切片的定性显微镜检查显示，rhEPO和CEPO均发挥了显著的锥体细胞保护作用。
我们的研究表明，在机械性脑外伤的体外模型中，10 IU/mL CEPO发挥的神经保护作用与rhEPO相当。由于CEPO缺乏造血作用且似乎具有延长的治疗时间窗，这种促红细胞生成素衍生物可能是治疗脑机械性损伤患者的一种令人兴奋的新药物工具。
