脊髓减压联合督脉电针对急性脊髓压迫损伤大鼠疗效影响的实验研究

Effects of Spinal Cord Decompression Surgery Combined with Du Meridian Electro-acupuncture for Treating Rats with Acute Spinal Cord Compression Injury

目的:探究脊髓减压联合督脉电针对持续脊髓压迫12h的急性脊髓损伤大鼠的治疗效果和可能的作用机理。方法:SPF级Wistar大鼠24只随机分为假手术组、督脉电针组、甲强龙组和模型组,每组6只,采用经寰枕间隙置入球囊导管加压造成脊髓压迫损伤的方法构建脊髓损伤模型。假手术组置入球囊导管后不加压,督脉电针组、甲强龙组及模型组加压12h后进行脊髓减压。督脉电针组取百会、大椎穴进行电针干预,连续波,频率2Hz,治疗时间15min,持续治疗14d;甲强龙组经大鼠尾静脉注射甲强龙进行冲击治疗;模型组不再进行干预。督脉电针干预14d后4组大鼠全部行脊髓损伤组织取材,采用BBB评分法对4组大鼠减压后0h,干预1d,3d,7d及14d后等5个时间点进行运动功能评价,ELISA法检测损伤组织血小板活化因子(Platelet-Activating Factor,PAF)的含量,Western blot法检测损伤组织中Caspase-3和Caspase-9的表达。结果:BBB评分:假手术组在5个时间点的评分均为(21±0.00)分,督脉电针组与甲强龙组各时间点评分相近,差异无统计学意义(P>0.05);督脉电针组与甲强龙组在干预1d,3d及7d后等时间点BBB评分较模型组高,差异有统计学意义(P<0.05);在干预14d时督脉电针组、甲强龙组及模型组BBB评分相近,差异无统计学意义(P>0.05)。ELISA法测PAF结果显示:模型组脊髓损伤组织中PAF浓度较假手术组、督脉电针组及甲强龙组高,差异有统计学意义(P<0.05);甲强龙组组织中PAF浓度较督脉电针组和假手术组高,差异有统计学意义(P<0.05);督脉电针组组织PAF浓度较假手术组高,差异有统计学意义(P<0.05)。Western blot法检测结果显示:Caspase-3和Caspase-9各组表达结果变化一致,模型组蛋白表达较假手术组、督脉电针组及甲强龙组高,差异有统计学意义(P<0.05);甲强龙组蛋白表达较督脉电针组和假手术组高,差异有统计学意义(P<0.05);督脉电针组蛋白表达较假手术组高,差异有统计学意义(P<0.05)。结论:脊髓减压联合电针通过降低脊髓损伤组织中PAF的含量与下调其Caspase9蛋白的表达来治疗急性脊髓压迫损伤,较单纯早期(<12h)减压可取得更好的临床效果。

Objective：To explore the therapeutic effect and underlying mechanism of spinal cord decompression surgery combined with Du meridian electro-acupuncture（DMEA）for treating the rats with acute spinal cord compression injury.Methods：All 24 SPF rats were randomly divided into sham-operated group,DMEA group,methylprednisolone group,and model group,with 6 rats in each group.The model of spinal cord injury was established by compressing the balloon catheter through the atlanto-occipital space,while the shamoperated group without compression.The spinal cord decompression surgery was using in DMEA group,methylprednisolone group,and model group 12 hafter modeling.The acupuncture point of Baihui and Dazhui were used in DMEA group,with continuous wave,frequency 2 Hz,and 15 min a day for 14 d.The rats in methylprednisolone group were received methylprednisolone impact therapy by tail vein injection.The model group did not receive any intervention.The injured spinal cord tissues were collected 14 dafter treatment.The function of hind limbs motor in each group at the five time points,0 h,1 d,3 d,7 d,and 14 dafter treating with DMEA were evaluated by BBB score method.The content of platelet activating factor（PAF）in injured spinal cord tissue were detected by ELISA method.And the expression of Caspase-3 and Caspase-9 in injured spinal cord tissue was detected by Western blot method.Results：The results of BBB score showed that the score of sham operation group was 21±0.00 at 5 time points,and there was no significant difference between DMEA group and methylprednisolone group at each time point（P〉0.05）.The score of BBB in methylprednisolone group was higher than that in model group at the time points of 1 d,3 d,7 d（P〈0.05）.The score of BBB in GVEA group,methylprednisolone group,and model group was similar at 14 d（P〉0.05）.The results of ELISA showed that the content of PAF in model group was higher than that in sham group,DMEA group and methylprednisolone group（P〈0.05）.The content of PAF in methylprednisolone group was higher than DMEA group and sham group.The content of PAF in DMEA group was higher than sham operation group（P〈0.05）.The results of Western blot showed that the expression of Caspase-3 and Caspase-9 in different groups were similar changed.The expression of the protein in model group were higher than that in Sham group,DMEA group and methylprednisolone group（P〈0.05）.The expression in methylprednisolone group was higher than that in DMEA group and Sham group（P〈0.05）.And the expression in DMEA group was higher than that in Sham group（P〈0.05）.Conclusion：Spinal cord decompression surgery combined with DMEA is more effective than surgery only for treating acute spinal cord compression injury though reducing the content of PAF and decreasing the expression of Caspase-9 protein.