Simon Florian Hans Peter, Erhart Philipp, Vcelar Brigitta, Scheuerle Angelika, Schelzig Hubert, Oberhuber Alexander
Department of Vascular and Endovascular Surgery, University of Düsseldorf, Düsseldorf, Germany.
Department of Vascular and Endovascular Surgery, University of Heidelberg, Heidelberg, Germany.
J Vasc Surg. 2016 Dec;64(6):1797-1804. doi: 10.1016/j.jvs.2015.10.011. Epub 2015 Nov 21.
This study examined effects and functional outcome of recombinant human erythropoietin (rhEPO) and carbamylated erythropoietin fusion protein (cEPO-FC) preconditioning in a rabbit model for spinal cord ischemia and resulting paraplegia. This model was chosen because only a small surgical effect is needed to cause paraplegia in rabbits, which facilitates postoperative observation of animals.
Anesthetized but spontaneously breathing New Zealand White rabbits randomly received cEPO-FC (50 μg/kg; n = 8), rhEPO (5000 IU/kg; n = 10), or vehicle (control; n = 10) 30 minutes before and after infrarenal aortic clamping. Ideal clamping time of 22 minutes was identified from preceding clamping tests (15-25 minutes). Postoperative observation time was 96 hours. Spinal cord function was assessed by neurologic evaluation of hind limb motor function every 12 hours using a modified Tarlov score. Spinal cord tissue damage was evaluated after 96 hours using hematoxylin and eosin, elastica van Gieson, Nissl, Masson-Goldner, and hemosiderin staining. Plasma levels of cell senescence markers stathmin, chitinase 1/3, elongation factor 1-α were determined.
Rabbits that received rhEPO showed significant improvement of spontaneous lower limb movements until 36 hours of reperfusion and improved histologic scores upon examination of the lumbar spinal cord compared with the control group. In contrast, cEPO-FC treatment showed comparable outcome to the control group concerning movements of the lower limbs and histology. Senescence markers were elevated in the control group, but not in the treatment groups, except for chitinase 3 in the rhEPO group. Only stathmin showed no significant effect. Markers for senescence might increase after acute ischemic injury. Attenuation of senescence markers might not come alone from improvement of the spinal cord.
Preconditioning with rhEPO attenuates ischemia/reperfusion injury of the spinal cord, whereas the carbamylated derivative (cEPO-FC) showed no positive effect on spinal cord function.
本研究在兔脊髓缺血及由此导致截瘫的模型中，检测重组人促红细胞生成素（rhEPO）和氨甲酰化促红细胞生成素融合蛋白（cEPO-FC）预处理的效果及功能转归。选择该模型是因为在兔中仅需较小的手术影响即可导致截瘫，这便于术后对动物进行观察。
麻醉但自主呼吸的新西兰白兔在肾下腹主动脉夹闭前后30分钟，随机接受cEPO-FC（50μg/kg；n = 8）、rhEPO（5000 IU/kg；n = 10）或溶剂（对照组；n = 10）。根据之前的夹闭试验（15 - 25分钟）确定理想的夹闭时间为22分钟。术后观察时间为96小时。每12小时使用改良的塔尔洛夫评分对后肢运动功能进行神经学评估，以评估脊髓功能。96小时后使用苏木精-伊红、弹性纤维van Gieson、尼氏、Masson-Goldner和含铁血黄素染色评估脊髓组织损伤。测定血浆中细胞衰老标志物Stathmin、几丁质酶1/3、延伸因子1-α的水平。
与对照组相比，接受rhEPO的兔在再灌注36小时前自发下肢运动有显著改善，且在检查腰段脊髓时组织学评分有所改善。相比之下，cEPO-FC治疗在下肢运动和组织学方面的结果与对照组相当。衰老标志物在对照组中升高，但在治疗组中未升高，rhEPO组中的几丁质酶3除外。只有Stathmin未显示出显著影响。衰老标志物可能在急性缺血性损伤后升高。衰老标志物的减弱可能并非仅源于脊髓功能的改善。
rhEPO预处理可减轻脊髓缺血/再灌注损伤，而氨甲酰化衍生物（cEPO-FC）对脊髓功能无积极影响。
