摘要
目的 观察IL-1β基因RNA干扰腺病毒载体对大鼠脊髓损伤（SCI）后运动功能恢复的影响,并探讨其可能机制. 方法 将48只健康雄性SD大鼠用压迫损伤法制成大鼠SCI模型,随机分为4组：正常对照组（Sham组）、脊髓损伤组（Vehicle组）、空载体组（Ad-GFP组）和IL-1β shRNA腺病毒组（Ad-shIL-1β组）.采用BBB评分评估大鼠后肢运动功能情况;荧光显微镜下观察GFP表达情况;ELISA法测定脊髓组织中IL-1β、TNF-α和IL-6蛋白表达水平;免疫荧光染色显示脊髓组织中NeuN阳性细胞的表达. 结果 SCI后1d,可观察到Ad-GFP组和Ad-shIL-1β组大鼠损伤部位脊髓组织有GFP表达;SCI后7d、14 d和21 d,Ad-shIL-1β组BBB评分（分别为8.17±1.17、10.17±0.98、11.33±0.82）明显高于Vehicle组（分别为4.00±0.89、5.67±1.03、6.17±1.17）和Ad-GFP组（分别为3.83±0.98、5.33±1.21、5.67±1.03）（P＜0.05）.SCI后21 d,Ad-shIL-1β组（115.17±25.77）的NeuN阳性细胞较Vehicle组（52.50±20.87）和Ad-GFP组（51.50±15.10）明显增加（P＜0.05）;SCI后1d,Ad-shIL-1β组IL-1β、TNF-α和IL-6蛋白表达（分别为138.83±7.96、143.38±10.20和120.43±9.79）较Vehicle组（分别为169.33±11.45、172.33±8.26、163.00±9.57）和Ad-GFP组（分别为172.83±10.85、167.48±8.19、159.48±10.98）明显降低（P＜0.05）. 结论 IL-1β表达水平降低能明显提高SCI后运动功能的恢复,对SCI具有保护作用.
Objective To investigate the possible protective effect of adenoviral vector expressing interleukin-1β （IL-1β） small hairpin RNA （shRNA） on spinal cord injury （SCI） and its mechanism in rats.Methods Forty-eight adult male Sprague-Dawley rats were randomly assigned to 4 groups including the Sham, the Vehicle,the Ad-GFP and the Ad-shIL-1β groups.SCI was induced by epidural compression.Motor function of hind limbs was evaluated by Basso-Beattie-Bresnahan （BBB） score, the expressions of green fluorescence in injured spinal cord tissue were observed by fluorescence microscope.Enzyme-linked immunosorbent assay （ELISA） and immunofluorescence were also performed.Results The expressions of green fluorescence in injured spinal cord tissue were observed in the Ad-GFP and Ad-shIL-1β groups one day after SCI.Significant functional improvement was observed in the Ad-shIL-1β group （8.17 ± 1.17, 10.17 ± 0.98 and 11.33 ± 0.82, respectively） compared to the Vehicle （4.00 ± 0.89, 5.67 ± 1.03 and 6.17 ± 1.17, respectively） and Ad-GFP （3.83 ± 0.98, 5.33 ± 1.21 and 5.67 ± 1.03, respectively） groups at 7, 14 and 21 days after SCI （P 〈 0.05）.Rats in the Ad-shIL-1β group had less neuronal loss 21 days after SCI.In addition, IL-1β downregulation significantly decreased IL-1β, tumor necrosis factor-or （TNF-α） and IL-6 levels （138.83 ± 7.96,143.38 ± 10.20 and 120.43 ± 9.79 in Ad-shIL-1β group;169.33 ± 11.45, 172.33 ± 8.26 and 163.00 ± 9.57 in Vehicle group;172.83 ± 10.85,167.48 ± 8.19 and 159.48 ± 10.98 in Ad-GFP group, respectively） one day after SCI （P 〈 0.05）.Conclusion This study demonstrated that the IL-1β downregulation may have potential therapeutic benefits for improving the outcomes after SCI.
出处
《中华显微外科杂志》
CSCD
北大核心
2015年第6期570-573,共4页
Chinese Journal of Microsurgery
基金
国家自然科学基金项目（81201403）
福建省教育厅科学基金项目（JB12093）
泉州市科技局科学基金项目（2012231）
