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山东大学学报(医学版) ›› 2010, Vol. 48 ›› Issue (4): 59-63.

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重组人促红细胞生成素抑制大鼠颈脊髓亚急性压迫性损伤中细胞凋亡的实验研究

宁斌1,张爱军2,贾堂宏1,宋宏亮1,郭舒亚1,龚维明1   

  1. 山东大学  1. 附属济南市中心医院骨外科, 济南 250013; 2. 低温医学实验室, 济南 250012
  • 收稿日期:2009-07-28 出版日期:2010-04-16 发布日期:2010-04-16
  • 通讯作者: 龚维明(1964- ),男,主任医师,主要从事脊柱疾患及骨创伤方面研究。 Email:wm-gong@hotmail.com
  • 作者简介:宁斌(1979- ),男,博士研究生,主治医师,主要从事脊柱疾患研究。 Email:ningbin@medmail.com.cn
  • 基金资助:

    济南市科技发展计划项目(200710)

Inhibitory effect of recombinant human erythropoietin on motor neuron apoptosis  during cervical chronic spinal cord compression in rats

NING Bin1, ZHANG Aijun2, JIA Tanghong1, SONG Hongliang1, GUO Shuya1, GONG Weiming1   

  1. 1. Department of Orthopedics, Affiliated Jinan Central Hospital of Shandong University, Jinan 250013, China;
    2.  Cryomedical Laboratory, Qilu Hospital of Shandong University, Jinan  250012, China
  • Received:2009-07-28 Online:2010-04-16 Published:2010-04-16

摘要:

目的  研究重组人促红细胞生成素(rhEPO)对大鼠颈脊髓亚急性压迫性损伤的治疗效果。方法  80只Wistar大鼠随机分为4组,假手术组(A组,n=5)仅行椎板切除,余大鼠用后路渐进性压迫法造成C5节段脊髓压迫性损伤。造模成功后,生理盐水对照组(B组,n=25)静脉给予生理盐水1mL/d,小剂量治疗组(C组,n=25)和大剂量治疗组(D组,n=25)分别静脉给予rhEPO 500u/(kg·d)、5000u/(kg·d);损伤后分别于3、7、14、21、28d用BBB评分评价四肢运动功能;免疫组化法检测受压颈髓节段EPO和EPO-R的分布及数量;各时间点处死模型,脊髓标本进行TUNEL染色和流式细胞仪定量检测细胞凋亡情况。结果   EPO-R随颈脊髓亚急性压迫性损伤渐重呈反应性增多,C、D组的各时间点BBB评分明显高于B组(P<0.01);TUNEL染色和流式细胞仪检测受压脊髓细胞凋亡结果显示:在21d时,rhEPO最能有效抑制脊髓运动神经元细胞的凋亡(P<0.01),小剂量rhEPO维持治疗给药剂量效果好。结论  颈脊髓亚急性压迫性损伤后给予rhEPO可减轻脊髓的运动神经元细胞的凋亡,促进脊髓运动功能恢复,且小剂量rhEPO维持治疗即可取得良好效果。

关键词: 亚急性颈脊髓压迫;重组人促红细胞生成素;细胞凋亡;大鼠,Wistar

Abstract:

Objective  To investigate effects of recombinant human erythropoietin (rhEPO) on chronic cervical spinal cord compression in rats. Methods  80 Wistar rats were randomly separated into 4 groups. Control rats (group A) received sham operations (n=5) and were executed by laminectomy. Experimental rats were subjected to compression injury in the C5 segment of the spinal cord by posterior progressive compression and subsequently subdivided into 3 groups. Group B (n=25) was treated with 1mL saline by IV. Groups C (n=25) and D (n=25) were treated with 500u/(kg·d) and 5000u/(kg·d) rhEPO by IV respectively. Limb motor function was scored using BBB standards 3, 7, 14, 21 and 28d after injury. The distribution and quantities of EPO and EPO-R in the compressed segment of the spinal cord were determined by immunobiochemical assay. Finally, apoptosis in the spinal cord was evaluated using TUNEL staining and flow cytometry at indicated time points. Results  The data indicated that expression of EPO-R increased significantly with aggravative level of chronic cervical spinal cord compression, as groups C and D exhibited higher BBB scores at all observed time points compared with group B(P<0.01). Using TUNEL staining and FCM, we observed that rhEPO could profoundly inhibit motor neuron apoptosis in the spinal cord (P<0.01) on day 21. Additionally, rhEPO treatment demonstrated a strong dose-dependent effect. Conclusions  rhEPO administration decreases motor neuron apoptosis in the spinal cord and improves spinal cord and motor functions following chronic cervical spinal cord compression. Moreover, continuing treatment with low doses of rhEPO shows a positive therapeutic effect.

Key words: Cervical spinal cord; Chronic spinal cord compression; recombinant human Erythropoietin (rhEPO); Apoptosis; Rats, Wistar

中图分类号: 

  • R683.2
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