Zhu Shuanglong, Duan Huiquan, Liu Yingfu, Li Guangzong, Liu Yingjie, Huang Mengqiang, Chen Xuyi, Xu Yunqiang
Graduate School, Tianjin Medical University, Tianjin, 300070, P.R.China;Department of Orthopedics, Tianjin Medical University General Hospital, Tianjin, 300052, P.R.China;Department of Brain, Affiliated Hospital, Logistics University of People's Armed Police Force, Tianjin, 300162, P.R.China.
Graduate School, Tianjin Medical University, Tianjin, 300070, P.R.China;Department of Orthopedics, Tianjin Medical University General Hospital, Tianjin, 300052, P.R.China.
Zhongguo Xiu Fu Chong Jian Wai Ke Za Zhi. 2017 Jul 15;31(7):825-829. doi: 10.7507/1002-1892.201702106.
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.
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 α (TNF-α) and interleukin 6 (IL-6) by ELISA, to detect the expressions of nuclear factor κB (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.
The BBB score and tiltboard experiment maximum angle were significantly higher in group A than groups B and C at each time point ( <0.05) and in group C than group B at 14, 21, and 28 days after operation ( <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 ( <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 ( <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, with hemorrhage, neutrophil infiltration, and nerve cell edema in the injured area; the neuronal cells were visible in the spinal cord of group C, with microglia mild hyperplasia, and the pathological changes were improved when compared with group B.
SSa has neuroprotective effects on acute spinal cord injury in rats by inhibiting NF-κB signaling pathway and AQP4 protein expression and reducing inflammation response and edema.
探讨柴胡皂苷a（SSa）对急性脊髓损伤大鼠免疫炎症水平的影响及其可能机制。
将72只体重220 - 250 g的Sprague Dawley大鼠随机分为假手术组（A组）、脊髓损伤组（B组）和SSa治疗组（C组），每组24只。B组和C组采用Allen法制备脊髓损伤模型；A组通过椎板切除术切断棘突和双侧椎板以暴露脊髓。损伤后即刻，C组大鼠腹腔注射10 mg/kg SSa，B组注射等体积生理盐水。术后24小时，每组取18只大鼠的脊髓组织，采用酶联免疫吸附测定（ELISA）法检测肿瘤坏死因子α（TNF-α）和白细胞介素6（IL-6）水平，采用蛋白质免疫印迹法检测核因子κB（NF-κB）P65、磷酸化核因子κB（NF-κB）P-P65和水通道蛋白4（AQP4）的表达，并通过苏木精-伊红（HE）染色观察脊髓形态。在损伤后1、3、7、14、21和28天，对6只大鼠的下肢运动功能进行Basso Beattie Bresnahan（BBB）评分和倾斜板实验评估。
各时间点A组的BBB评分和倾斜板实验最大角度均显著高于B组和C组（P<0.05），术后14、21和28天C组高于B组（P<0.05）。ELISA检测显示，A组TNF-α和IL-6浓度显著低于B组和C组，C组低于B组（P<0.05）。蛋白质免疫印迹结果显示，A组NF-κB P65、NF-κB P-P65和AQP4的蛋白表达水平显著低于B组和C组，C组低于B组（P<0.05）。HE染色显示，A组脊髓神经元正常，无明显病变；B组损伤区域可见神经元细胞，损伤区域有出血、中性粒细胞浸润和神经细胞水肿；C组脊髓可见神经元细胞，小胶质细胞轻度增生，与B组相比病理变化有所改善。
SSa可通过抑制NF-κB信号通路和AQP4蛋白表达，减轻炎症反应和水肿，对大鼠急性脊髓损伤具有神经保护作用。
