三羟乙基芦丁对大鼠颈脊髓损伤后神经功能恢复的影响

Effects of trihydroxyethyl rutin on the recovery of nerve function after cervical spinal cord injury in rats

中文关键词:  三羟乙基芦丁  脊髓损伤  行为学  组织学  微血管

英文关键词:Trihydroxyethyl rutin  Spinal cord injury  Behavior  Histology  Microvascular

中文摘要:

  【摘要】 目的：探讨三羟乙基芦丁对大鼠颈脊髓损伤的神经保护作用。方法：将成年雄性SD大鼠44只随机分为4组，假手术组（Shame组，n=8）；损伤组（Injury组，n=12）；三羟乙基芦丁50mg/kg治疗组（T50组，1次/日，腹腔注射，n=12）；三羟乙基芦丁100mg/kg治疗组（T100组，1次/日，腹腔注射，n=12），后三组均采用C5脊髓半侧挫伤大鼠模型（挫伤深度为1.2mm）。分别于术前及术后3d、1周、2周、4周、6周、8周、10周、12周评价各组大鼠前肢运动功能评分（forelimbs locomotor scores，FLS）、梳理实验等行为学变化。12周时各组大鼠随机选取1只进行硫酸钡-明胶灌注剂进行颈脊髓微血管灌注，4℃过夜后显微镜下取颈脊髓组织，免疫组织化学染色观测颈脊髓微血管改变，脊髓组织连续冰冻切片铬花青EC染色统计各组损伤情况。结果：术后3d时，Injury组、T50组及T100组的FLS分别为11.33±0.71分、11.55±0.81分和12.36±1.15分，与Shame组（17.0分）相比均显著下降，随着时间的推移，FLS均逐渐改善，T50组和T100组于术后8周时恢复正常，Injury组于术后10周时恢复正常。梳理实验结果与FLS相似，至12周时T100组恢复正常（5.0分），T50组为4.92±0.08分，与Shame组比较无显著性差异，Injury组为4.17±0.11分，与Shame组比较仍有显著性差异（P＜0.05）。术后12周时，Injury组微血管数量明显减少，T50组也明显减少，但优于Injury组，T100组的微血管数量减少不明显；免疫组化染色显示，12周时Injury组、T50组及T100组微血管密度分别为23.50±1.75、31.33±1.36和33.00±1.27，均较Shame组明显减少，T50组、T100组与Injury组比较有显著性差异（P＜0.05）。EC染色结果发现，T50组和T100组损伤侧的总面积（包括空洞面积）明显高于Injury组；Injury组损伤中心的空洞面积最大，而T50组和T100组损伤中心空洞面积显著小于Injury组；T50组和T100组损伤侧残余灰质面积均明显多于Injury组。结论：三羟乙基芦丁通过保护颈脊髓微血管结构，减轻脊髓结构破坏，对大鼠颈脊髓损伤具有较好的神经保护作用。

英文摘要:

  【Abstract】 Objectives: To investigate the neuroprotective effect of trihydroxyethrutin on cervical spinal cord injury in rats. Methods: 44 adult male SD rats were randomly divided into 4 groups, including the shame group(n=8), injury group(n=12), T50 group with 50mg/kg trihydroxyethyl rutin treatment(1 time/day, intraperitoneal injection, n=12), and T100 group with 100mg/kg trihydroxyethyl rutin treatment(1 time/day, intraperitoneal injection, n=12). The last three groups were all treated with C5 spinal cord hemilateral contusion rat model(contusion depth was 1.2mm). The behavioral changes such as forelimb locomotor scores(FLS) and combing experiment were evaluated at pre-operation and 3d, 1 week, 2 weeks, 4 weeks, 6 weeks, 8 weeks, 10 weeks and 12 weeks after operation. At 12 weeks, 1 rat in each group was randomly selected to inject with barium-sulfate gelatin for cervical spinal cord microvascular perfusion. After 4℃ overnight, cervical spinal cord tissue was collected under the microscope. Changes in cervical spinal cord microvascular were observed by immunohistochemical staining. Results: At 3 days after the surgery, the FLS of the injury group, the T50 group and the T100 group were 11.33±0.71, 11.55±0.81 and 12.36±1.15 points, respectively. At 8 weeks after the surgery, compared with the shame group(17.0 points), the FLS of the injury group, the T50 group and the T100 group returned to normal. And at 10 weeks after the surgery, the injury group returned to normal. The results of the grooming experiment were similar to those of FLS, with the T100 group returning to normal(5.0 points) at 12 weeks, the T50 group returning to normal at 4.92±0.08 points, and no significant differences between the shame group and the injury group(4.17±0.11 points)(P<0.05). At 12 weeks after surgery, the number of microvessels in the injury group was significantly reduced, and so was that in the T50 group, while it was better than that in the injury group. The number of microvessels in the T100 group was not significantly reduced. Immunohistochemical staining showed that at 12 weeks, the microvascular density of the injury group, the T50 group and the T100 group were 23.50±1.75, 31.33±1.36 and 33.00±1.27, respectively, which were significantly lower than those of the shame group, and there were significant differences between the T50 group and the T100 group and the injury group(P<0.05). The EC staining results showed that the T50 group and T100 group had significantly higher total area of injury side(including cavity area) than the injury group. The injury group had the largest hole area, while the T50 and T100 groups had significantly smaller holes than the injury group. The residual gray matter area of T50 group and T100 group was significantly higher than that of the injury group. Conclusions: Trihydroxyethyl rutin provides neuroprotection in rats with cervical spinal cord injury, by protecting the microvascular structure of cervical spinal cord and alleviating the damage of spinal cord structure.