重组人红细胞生成素治疗大鼠急性脊髓损伤的用药时间窗探讨

Effect of recombinant human erythropietin administrated at different time point on neurofunction recovery after acute SCI：an experimental study on rat model

中文关键词:  急性脊髓损伤  重组人红细胞生成素  治疗  细胞凋亡  大鼠

英文关键词:Acute spinal cord injury  Recombinant human erythropoietin  Pharmacotherapy  Apoptosis  Ｒat

中文摘要:

  【摘要】 目的：探讨腹腔注射重组人红细胞生成素（rHuEPO）治疗大鼠急性脊髓损伤（ASCI）的用药时间窗。方法：采用显微血管夹夹伤雌性SD大鼠T10脊髓建立脊髓急性损伤模型，将造模成功后的40只大鼠随机分为A、B、C、D、E组，每组8只。A、B、C、D组分别于损伤后即刻、1h、3h和6h腹腔注射rHuEPO 5000IU/kg，E组损伤后立即腹腔注射等量生理盐水，作为对照组。各组大鼠于术后1d、3d、5d、7d进行BBB运动学评分，并于术后3d、7d用TUNEL染色和caspase-3免疫组化染色法检测各组大鼠脊髓神经细胞凋亡情况。结果：术后1d、3d各组大鼠ＢＢＢ评分无统计学差异（P>0.05）；5d时A、B、C三组ＢＢＢ评分均高于D、E两组（P<0.01），A、B、C三组间与D、E组间无统计学差异（P>0.05）；7d时D组BBB评分高于E组（P<0.01），但低于A、B、C三组（P<0.01），而A、B、C三组间无统计学差异（P>0.05）。损伤后3d、7d，A~D组的TUNEL、caspase-3染色阳性细胞均高于E组，且损伤后7d时D组TUNEL染色阳性细胞数高于A、B、C三组（P<0.05），A、B、C三组间无统计学差异；损伤后3d，Ｄ组caspase-3染色细胞阳性率高于A、B、C三组（P<0.05），A、B、C三组间无统计学差异（P>0.05），损伤后7d，A~D组间无统计学差异（P>0.05）。结论：大鼠ASCI后3h内给予rHuEPO可以显著改善运动功能恢复情况，并抑制伤后脊髓神经细胞凋亡现象的发生；伤后6h给药效果较差。

英文摘要:

  【Abstract】 Objective：To identify the effect of recombinant human erythropoietin（rHuEPO） administrated at different time point on neurofunction recovery after SCI.Method：An acute spinal cord injury（ASCI） animal model was induced by clip of T10 spinal cord.After ASCI，forty female rats were randomly divided into five groups with eight rats in each group.Groups A，B，C and D received 5000IU/kg of rHuEPO by intraperitoneal injection at 0 min，1 hour，3 hours and 6 hours after ASCI respectively，with single dose for each group. Group E received the same dose of saline solution immediately after ASCI.BBB motor function score was recorded at 4 post-injury time points（1，3，5，7 days），and TUNEL staining and anti-active caspase-3 staining was used to evaluate the cell apoptosis at 3，7 days after injury.Result：BBB score at 1 day and 3 days among each group showed no diffference（P>0.05），while at 5 days after injury，BBB score of group A，B and C were higher than that of group D and E（P<0.01）.No significant intergroup difference were noted among group A，B，C and group D，E（P>0.05）.At 7 days after injury，BBB score of group D was higher than group E（P<0.01），while lower than group A，B and C（P<0.01），and no difference were noted among group A，B and C（P>0.05）.At 3 and 7 days after injury，the number of TUNEL-positive cells in group A，B and C was significantly less than those in group E（P<0.01），and TUNEL-positive cells in group D were more than those in group A，B and C at 7 days after injury（P<0.05），no difference were noted among group A，B and C（P>0.05）.Caspase-3 positive cells in group D were more than those in group A，B and C at 3 days after injury（P<0.05），while no difference were noted among group A，B and C（P>0.05）.At 7 days after injury，no difference were noted among group A，B，C and D（P>0.05） with regard to caspase-3 positive cells（P<0.05）.Conclusion：Administration of rHuEPO within 3 hours after ASCI can significantly improve motor function and inhibit neuron apoptosis，while this effect remains fair when administered over 6 hours after ASCI.