西维来司他钠在大鼠急性脊髓损伤中的保护作用

Protection of Sivelestat sodium on acute spinal cord injury in rats

目的探讨脊髓损伤后早期使用西维来司他钠(Sivelestat sodium)抑制细胞凋亡,对大鼠脊髓损伤的保护作用及机制。方法将60只SD大鼠随机分成5组,每组12只,Allen's脊髓损伤模型后,立即腹腔注射Sivelestat sodium 1.6 mg/kg,4.8 mg/kg,10 mg/kg,50 mg/kg,并连续7 d(1次/d),对照组给等量生理盐水,并采用脊髓运动功能评分(BBB scale)对大鼠进行双下肢神经功能评分。原位末端标记法(TUNEL)观察脊髓损伤后脊髓神经细胞凋亡情况。酶联免疫吸附法(ELASA)检测炎症因子。免疫印迹法检测甘油醛-3-磷酸脱氢_E3泛素连接酶-1(GAPDH-Siah1)细胞凋亡序列。结果以BBB标准评价大鼠的双下肢运动功能,治疗组的下肢活动功能明显好于对照组。4.8 mg/kg和10 mg/kg剂量组双下肢神经功能学评分与对照组比较具有明显差异(P<0.01);50 mg/kg剂量组未见明显保护作用,因此未加入统计结果。TUNEL染色显示,sivelestat sodium明显减少了脊髓损伤后神经细胞凋亡,抑制脊髓损伤后炎症因子表达。sivelestat sodium显著抑制GAPDH-Siah1凋亡序列的表达。结论 sivelestat sodium通过抑制GAPDH-Siah1凋亡序列,减少脊髓损伤后脊髓神经细胞的凋亡,从而改善脊髓损伤大鼠后肢运动神经功能。

Objective The aim of this study was to investigate wnether Sivelestat sodium treatment could protect the spinal cord against experimental spinal cord injury through anti-apoptosis.Methods A total of 60 male SD rats were randomly divided into five groups,control group,1.6 mg/kg group,4.8 mg/kg group,10 rng/kg group,and 50 mg/kg group （n =12）.After the model of moderate spinal cord injury was established according to the Allen＇s method,Sivelestat sodium （1.6 mg/kg,4.8 mg/kg,10 rng/kg,50 mg/kg）or a saline control was administered by intraperitoneal injection for 7 d after spinal cord injury （SCI）.Locomotor recovery was tested with the Basso Beattie Bresnahan locomotor rating scale（BBB scale）at 1 d,3 d,7 d,14 d,21 d after injury,respectively.Neuronal apoptosis were observed by TdT-mediated UTP nick end labeling （TUNEL dyeing） in different groups.In the spinal cord tissues,IL-1β,TNF-α,and NFKB were analyzed by enzyme-linked immunosorbent assay （ELASA） and apoptosis sequence of glyceraldehyde-3-phosphate dehydrogenase-seven in absentia homolog 1 （GAPDH-Siah1） were detected by Western blotting.Results Lower extremity function in the treatment groups of 4.8 mg/kg and 10 mg/kg was significantly improved compared with control group （P ＜ 0.01）.There were significant differences between treatment and control groups in HE staining and TUNEL dyeing.The levels of pro-inflammation factors,such as,interleukin 1β （IL-1β）,tumor necrosis factor （TNF-α）,nuclear factor-k-gene binding （NFKB） were lower in treatment group than those in control group （P ＜0.01）.Western blotting showed that the expression of GAPDH-Siah1 was inhibited in treatment group obviously （P ＜0.01）.Conclusion Sivelestat sodium reduces the neural apoptosis after spinal cord injury by inhibiting the expression of GAPDH-Siah1 and accordingly improves the locomotor function recovery.