柴胡皂苷a对大鼠急性脊髓损伤的神经保护作用与机制研究  

Neuroprotective effects and mechanism of saikosaponin A on acute spinal cord injury in rats

作　　者：朱双龙[1,2,3] 段会全[1,2] 刘英富[4] 李光宗[5] 刘迎节[1,2] 黄梦强 陈旭义[3] 徐云强[1,2] 

机构地区：[1]天津医科大学研究生院,天津300070 [2]天津医科大学总医院骨科,天津300052 [3]武警后勤学院附属医院脑科中心,天津300162 [4]武警后勤学院细胞生物学教研室,天津300309 [5]武警后勤学院附属医院救援研究所,天津300162

出　　处：《中国修复重建外科杂志》2017年第7期825-829,共5页Chinese Journal of Reparative and Reconstructive Surgery

基　　金：国家科技重点研发计划资助项目(2016YFC1101500);国家自然科学基金面上项目(11672332);国家自然科学基金青年项目(81101362)~~

摘　　要：目的探讨柴胡皂苷a(saikosaponin a,SSa)对大鼠急性脊髓损伤早期机体免疫炎性水平的影响,并研究其可能存在的影响机制。方法取健康成年雌性SD大鼠72只,体质量220~250 g,随机分为假手术组(A组)、脊髓损伤组(B组)、SSa处理组(C组),每组24只。A组仅咬除棘突和椎板暴露脊髓;B、C组利用改良Allen重物打击法制作急性脊髓损伤模型,造模后即刻C组给予大鼠腹腔注射10 mg/kg SSa,B组注入等量生理盐水。术后24 h每组处死18只大鼠并取出脊髓组织,采用ELISA法检测TNF-α和IL-6浓度,Western blot检测NF-κB P65、NF-κB P-P65和水通道蛋白4(aquaporin 4,AQP4)蛋白表达水平,HE染色观察脊髓组织形态;每组其余6只大鼠分别于术后1、3、7、14、21、28 d采用BBB评分和斜板实验评价大鼠双下肢运动恢复情况。结果术后各时间点A组BBB评分和斜板实验最大角度均显著高于B、C组(P<0.05),术后14、21、28 d C组上述指标显著高于B组(P<0.05)。ELISA检测示,A组TNF-α、IL-6浓度显著低于B、C组,C组显著低于B组,差异均有统计学意义(P<0.05)。Western blot检测示,A组NF-κB P65、NF-κB P-P65和AQP4蛋白表达水平显著低于B、C组,C组显著低于B组,差异均有统计学意义(P<0.05)。HE染色示,A组脊髓中神经细胞正常,无明显病变;B组脊髓中损伤部位可见神经元细胞,损伤处有出血、中性粒细胞浸润、神经细胞水肿;C组脊髓中可见神经元细胞,小胶质细胞轻度增生,病变较B组好转。结论 SSa通过抑制NF-κB信号通路和AQP4蛋白的表达,减轻急性脊髓损伤后机体炎性反应和组织水肿,从而具有一定保护神经的功能。Objective To investigate the effect of saikosaponin a （SSa） on the levels of immune inflammation in rats with acute spinal cord injury and its possible mechanism. Methods Seventy-two Sprague Dawley rats （weighing, 220-250 g） were randomly divided into sham operation group （group A）, spinal cord injury group （group B）, and SSa treatment group （group C） respectively, 24 rats in each group, The spinal cord injury model was induced by using the Allen＇s method in groups B and C; the spinous process and vertebral plate at both sides were cut off by lamina excision to expose the spinal cord in group A. The rats were given intraperitoneal injection of 10 mg/kg SSa in group C and equal volume of normal saline in group B at immediate after injury. The spinal cord tissue was harvested from 18 rats of each group at 24 hours after operation to measure the levels of tumor necrosis factor ct （TNF-α） and interleukin 6 （IL-6） by ELISA, to detect the expressions of nuclear factor KB （NF-κB） P65, NF-κB P-P65, and aquaporin 4 （AQP4） by Western blot and to observe the morphology of spinal cord by HE staining. The motor function of the lower limbs was evaluated by BBB score and tiltboard experiment in 6 rats at 1, 3, 7, 14, 21, and 28 days after injury. Results The BBB score and tiltboard experiment maximum angle were significantly higher in group A than groups B and C at each time point （P〈0.05） and in group C than group B at 14, 21, and 28 days after operation （P〈0.05）. ELISA test showed that the concentrations of TNF-α and IL-6 were significantly lower in group A than groups B and C, and in group C than group B （P〈0.05）. Western blot results showed that the protein expression levels of NF-κB P65, NF-κB P-P65, and AQP4 were significantly lower in group A than groups B and C, and in group C than group B （P〈0.05）. HE staining demonstrated normal neurons of the spinal cord and no obvious lesion in group A; neuronal cells were observed in the injured area of group B, wi

关 键 词：柴胡皂苷A 脊髓损伤 炎性反应 水通道蛋白4 神经保护 大鼠 

分 类 号：R651.2[医药卫生—外科学]