甲泼尼龙琥珀酸钠鞘内注射治疗急性脊髓损伤的实验研究

Effects of intrathecal injection of methylprednisolone sodium succinate in acute spinal cord injury rabbits

目的研究甲泼尼龙琥珀酸钠（MPss）鞘内注射治疗新西兰兔急性脊髓损伤的疗效及神经元凋亡情况。方法选取健康新西兰兔72只，随机分为急性脊髓损伤组（SCI组）及假手术组（SHAM组），每组分成1～6亚组，分别给予生理盐水、鞘内注射MPss、静脉注射MPss及联合应用鞘内及静脉注射MPss治疗。术后每天行TARLOV下肢运动功能评分，7d处死实验动物，取脊髓损伤、假手术部位及鞘内注射部位神经组织标本，予以HE染色和TUNEL染色。不同时间点TARLOV评分比较应用两因素重复测量资料方差分析，组间应用单因素方差分析进行统计学处理。结果术后7d时，SCI组鞘内注射MPss3．0mg／kg、6．0mg／kg与传统静脉注射治疗的TARLOV评分差异无统计学意义（P〉0．05），且均高于生理盐水对照组（F=4．762，P〈0．05）；SCI组鞘内注射MPss6．0mg／kg的脊髓损伤部位淋巴细胞浸润评分为（1．33±0．21）分，与生理盐水对照组的（2．67± O．21）分比较差异有统计学意义（F=5．793，P〈0．05），与传统静脉注射治疗差异无统计学意义（P〉0．05）。SHAM组鞘内注射MPss6．0mg／kg的鞘内注射部位淋巴细胞浸润评分为（2．50±O．55）分，与生理盐水对照组的（0．504-0．55）分比较差异有统计学意义（F：17．333，P〈0．05）；SC!组TUNEL染色提示鞘内注射6．0mg／kg的凋亡细胞计数[（6．3±1．5）个]与生理盐水对照组[（20．3±2．2）个]比较差异有统计学意义（F=71．279，P〈0．05）。结论鞘内注射MPss可以改善新西兰兔急性脊髓损伤后下肢运动功能，取得与传统静脉冲击治疗相似的疗效，对抗神经细胞凋亡，改善脊髓损伤预后。

Objective To investigate the effects and apoptosis of intrathecal injection of Methylprednisolone Sodium Succinate （MPss） for acute spinal cord injury （SCI） in New Zealand rabbits. Methods Seventy-two healthy New Zealand rabbits were used for the procedure and were randomly divided into two groups ： SCI group and SHAM group, which was both divided into 6 subgroups, such as the vehicle group, the MPss intrathecal injection groups （1.5 mg/kg, 3.0 mg/kg, 6. 0 mg/kg group ）, the MPss intravenous injection group and the combined injection group. TARLOV score was tested daily to evaluate the motor function. The rabbits were sacrificed 7 days after the surgery and the thoracic spinal cord sections and the sacral sections where MPss was injected were harvested for HE and TUNEL staining. Two-Factors Repeated Measures analysis of variance for TARLOV scores tested at various times and One-Way ANOVA analysis of variance for data between groups were used. Results Seven days after surgery in SCI group, there was no statistical difference between the TARLOV scores of intrathecal injection of MPss 3.0 mg/kg group, 6. 0 mg/kg group and MPss intravenous injection group （ P 〉 0. 05 ）, which were all better than the vehicle group （ F = 4. 762, P 〈 0. 05 ）. Referring to the lymphocyte infiltration at the injury site in SCI group, there was statistical difference between MPss intrathecal injection 6. 0 mg/kg group （ 1.33 ± 0. 21 ） and the vehicle group（ 2. 67 ± 0. 21 ） （F = 5. 793 ,P 〈 0. 05 ） and no statistical difference between intrathecal injection of MPss 6. 0 mg/kg group and MPss intravenous injection group （P 〉 0. 05 ）. As for the lymphocyte infiltration at the intrathecal injection site in SHAM group, there was statistical difference between MPssintrathecal injection 6. 0 mg/kg group（2. 50 ±0. 55） and the vehicle group（0. 50 ±0. 55） （F = 17. 333 ,P 〈 0. 05 ）. TUNEL staining in SCI group showed statistical difference between MPss intrathecal injection 6. 0 mg/kg group （6. 3 ± 1.5 ） and the vehicle group （ 20. 3 ± 2. 2 ） （ F = 71. 279, P 〈 0. 05 ）. Conclusions Intrathecal injection of MPss can improve the functional recovery of lower limb and decrease apoptosis of neuron cells, which can provide same effects as the traditional intravenous injection of MPss in New Zealand rabbits.