摘要
研究大黄素对大鼠急性脊髓损伤后氧化应激和炎症反应的影响,探讨大黄素对急性脊髓损伤后神经元的保护机制。以Allen’s法制备脊髓损伤模型。将195只SD大鼠随机分为假手术组（A组）、模型组（B组）、大黄素20 mg·kg-1组（C组）、大黄素40 mg·kg-1组（D组）、大黄素80 mg·kg-1组（E组）,术后3,7,14,28 d予BBB评分,斜板实验观察大鼠运动功能恢复。术后7 d,甲苯胺蓝染色观察神经元尼氏小体,电镜观察脊髓神经纤维改变;Western blot法检测Nrf2,HO-1,NQO1,GFAP,NF-κB蛋白表达;ELISA检测TNF-α,IL-1β,IL-6含量;免疫荧光检测GFAP,NG2,ED-1表达。BBB评分与斜板实验显示,术后7,14,28 d,C,D,E组较B组明显升高,差异均具有统计学意义（P〈0.05）;术后7 d,B,C组尼氏小体开始融合,D,E组较B,C组融合明显减轻;电镜显示,B,C组脊髓神经纤维出现明显脱髓鞘改变,D,E组髓鞘变性较B,C组减轻;Nrf2,HO-1,GFAP,NF-κB蛋白表达C,D,E组与B组比较,NQO1蛋白表达D,E组与B组比较,差异均有统计学意义（P〈0.05）。C,D,E组TNF-α,IL-6含量较B组,D,E组IL-1β含量较B组显著降低,差异均有统计学意义（P〈0.05）。C,D,E组GFAP,NG2表达较B组增加,D,E组更明显;C,D,E组ED-1表达较B组显著降低。大黄素对脊髓急性损伤后神经细胞具有保护作用,其机制可能与激活Nrf2-ARE通路,降低NF-κB,ED-1,TNF-α,IL-1β,IL-6表达,促进GFAP,NG2表达有关。
This paper was aimed to investigate the effects of emodin on oxidative stress and inflammatory response in rats with acute spinal cord injury（ SCI）,and to explore the protective mechanism of emodin on neurons after SCI. Rat SCI models were established using a modified Allen’s method. One hundred and ninety five Sprague-Dawley rats were randomly divded into sham（ group A）,model（ group B）,emodin group of 20 mg·kg-1（ group C）,emodin group of 40 mg·kg-1（ group D）,emodin group of 80 mg·kg-1（ group E）. Functional recovery was evaluated using the Basso-Beattie-Bresnahan（ BBB） scale and inclined plate test on the 3 rd,7 th,14 th and 28 th day. On the 7 th day after SCI,neuron nylon body was observed with toluidine blue staining. The changes of myelinated nerve fibers were observed by transmission electron microscope. Nrf2,HO-1,NQO1,GFAP,NF-κB protein were detected by Western blot.The content of TNF-α,IL-1β,IL-6 were detected by ELISA. Immunofluorescence staining was performed to observe the expression of the GFAP,NG2 and ED-1. The BBB and inclined plate scores of group C,D and E were higher than group B on the 7 th,14 th,28 th day,the difference was statistically significant（ P 〈 0. 05）. On the 7 th day,Nylon Body of group B and C started to fuse,the fusion of group D and E were significantly alleviated than group B and C. Transmission electron microscope showed that the changes of demyelination were obvious in group B and C,group D and E were significantly improved than group B and C. Western blot showed that Nrf2,HO-1,GFAP,NF-κB protein expression,group C,D,E compared with group B,NQO1 protein expression,group D,E compared with group B,the difference was statistically significant（ P 〈 0. 05）. ELISA showed that the content of TNF-α,IL-6,group C,D,E compared with group B,the content of IL-1β,group D,E compared with group B,the difference was statistically significant（ P 〈0. 05）; Immunofluorescence showed that the expressions of GFAP and NG2 in group C,D and E were higher than that in group B,and the group D and E were more obvious. The expression of ED-1 in group C,D,E were decreased significantly compared with group B.Emodin has protective effect on neurons after SCI. The mechanism may connect with activting Nrf2-ARE pathway,reducing the expression of NF-κB,ED-1,TNF-α,IL-1β,IL-6,and promoting the expression of GFAP and NG2.
作者
曾欢欢
黄英如
李子健
汪一
张松
ZENG Huan-huan;HUANG Ying-ru;LI Zi-jian;WANG Yi;ZHANG Song(Department of Acupuncture and Orthopaedics of Traditional Chinese Medicine College, Chongqing Medical University, Choagqing 400016, China;Chongqing Key Laboratory of Traditional Chinese Medicine for Prevention and Cure of Metabolic Diseases, Chongqing 400016, China)
出处
《中国中药杂志》
CAS
CSCD
北大核心
2018年第9期1886-1893,共8页
China Journal of Chinese Materia Medica
基金
国家自然科学基金面上项目(81373668,81674002)
关键词
脊髓损伤
大黄素
氧化应激
炎症
胶质纤维酸性蛋白
神经元胶原抗原2
spinal cord injury
emodin
oxidative stress
inflammation
glial fibrillary acid protein
neuronglialantigen-2
作者简介
曾欢欢，硕士研究生，E-mail：719853539@qq．com;[通信作者]黄英如，副教授，研究方向为脊髓与周围神经损伤，E-mail：hyr12678@126．com
