摘要
目的探究抑制炎性介质肿瘤坏死因子-α(TNF-α)在脊髓损伤后神经修复中的作用,并揭示其具体作用机制。方法实验研究起始时间为2023年3月至2024年6月,通过动脉瘤夹压迫型损伤法制备脊髓损伤大鼠模型,采用随机数表法将45只雌性SD大鼠分为3组,分别为假手术组、脊髓损伤组、TNF-α抑制剂组,每组大鼠15只。除假手术组外,其余组大鼠均通过动脉瘤夹压迫型损伤法构建脊髓损伤大鼠模型,TNF-α抑制剂组大鼠鞘内注射10 mg/kg XPro®1595,其余组大鼠注射等体积生理盐水。采用Basso-Beattie-Bresnahan评分法和后肢运动轨迹法检测后肢运动恢复能力;免疫荧光染色分析脊髓损伤后大鼠的神经再生;Nissl染色评估各组大鼠神经轴突的再生;蛋白质印迹法(Western blot)检测炎症相关通路的蛋白水平。结果实验结果显示,TNF-α抑制剂组大鼠BBB评分(25.26±4.84)明显高于脊髓损伤组(17.52±3.98),低于假手术组(32.23±3.21),差异有统计学意义(t=4.784、4.648,P<0.000)。TNF-α抑制剂组大鼠后肢运动功能中的步幅长度[(12.48±2.52)cm]明显低于脊髓损伤组[(25.68±5.36)cm],高于假手术组[(4.73±0.84)cm],差异有统计学意义(t=8.631、11.299,P<0.000)。TNF-α抑制剂组大鼠神经轴突数量(182.45±24.74)明显高于脊髓损伤组(88.36±12.20),低于假手术组(249.62±32.65),差异有统计学意义(t=10.786、5.185,P<0.001)。TNF-α抑制剂组大鼠损伤区域的脊髓组织尼氏小体数量(45.78±7.82)明显低于脊髓损伤组(80.46±12.49),高于假手术组(18.62±3.03),差异有统计学意义(t=7.442、10.241,P<0.01)。TNF-α抑制剂组TNF-α、IL-1β、IL-6、NF-κB p65蛋白水平(3.86±1.24、5.74±1.88、5.26±1.46)明显低于于脊髓损伤组(7.55±2.61、11.87±3.25、9.53±2.08),差异有统计学意义(t=4.867、4.769、4.727,P<0.05)。结论抑制炎性介质TNF-α对脊髓损伤后大鼠具有明显的神经修复作用,其机制可能与抑制炎症相关信号通路有关。
Objective To explore the role of inflammatory mediators tumor necrosis factor-α(TNF-α)in nerve repair after spinal cord injury(SCI)and reveal its specific mechanism.Methods The experimental study started from March 2023 to June 2024.The rat model of spinal cord injury was prepared by aneurysm clamp compression injury method.45 female SD rats were randomly divided into 3 groups by random number table method,which were as follows:Sham operation group,spinal cord injury group and TNF-αinhibitor group included 15 rats in each group.Except the sham operation group,the spinal cord injury rat model was constructed by aneurysm clamp compression injury method.The TNF-αinhibitor group received intravaginal injection of 10 mg/kg XPro®1595,and the other group received intravaginal injection of equal volume normal saline.Basso-Beattie-Bresnahan scoring method and hind limb motion trajectory method were used to detect hind limb motion recovery ability.Nerve regeneration in rats after SCI was analyzed by immunofluorescence staining.The regeneration of nerve axons in each group was evaluated by Nissl staining.Western blotting was used to detect protein levels in inflammatory pathways.Results The results showed that the BBB score of TNF-αinhibitor group(25.26±4.84)was significantly higher than that of spinal cord injury group(17.52±3.98),and lower than that of sham operation group(32.23±3.21),with statistical significance(t=4.784,4.648,P<0.01).The stride length of hind limb motor function of rats in TNF-αinhibitor group[(12.48±2.52)cm]was significantly lower than that in spinal cord injury group[(25.68±5.36)cm],and higher than that in sham operation group[(4.73±0.84)cm],the difference was statistically significant(t=8.631,11.299,P<0.01).The number of neuronal axons in TNF-αinhibitor group(182.45±24.74)was significantly higher than that in spinal cord injury group(88.36±12.20)and lower than that in sham operation group(249.62±32.65),the difference was statistically significant(t=10.786,5.185,P<0.001).The number of Nishi bodies in the injured area of the spinal cord in the TNF-αinhibitor group(45.78±7.82)was significantly lower than that in the injured group(80.46±12.49),and higher than that in the sham operation group(18.62±3.03),with statistical significance(t=7.442,10.241,P<0.01).The protein levels of TNF-α,IL-1β,IL-6 and NF-κB p65 in TNF-αinhibitor group(3.86±1.24,5.74±1.88,5.26±1.46)were significantly lower than those in spinal cord injury group(7.55±2.61,11.87±3.25,9.53±2.08).The difference was statistically significant(t=4.867,4.769,4.727,P<0.05).Conclusion Inhibition of inflammatory mediators TNF-αcan significantly repair nerves after SCI in rats,and the mechanism may be related to inhibition of inflammatory signaling pathways.
作者
王冰一
陈光
高强
蔡腾
刘涛
Wang Bingyi;Chen Guang;Gao Qiang;Cai Teng;Liu Tao(Department of Orthopaedic,Henan Provincial People’s Hospital,Zhengzhou 450003,China)
出处
《中华实验外科杂志》
2025年第4期692-695,共4页
Chinese Journal of Experimental Surgery
基金
河南省重点研发专项项目(241111313400)。
关键词
炎症
肿瘤坏死因子-Α
脊髓损伤
神经修复
Inflammation
Tumor necrosis factor-α
Spinal cord injury
Nerve repair
