大鼠脊髓损伤早期CNTF与NT-4表达变化及其相互关系  

The Expression Changes of CNTF and NT-4 in Rats with Early Spinal Cord Injury and Their Correlation

作　　者：牛英杰[1] 吴潘娜 胡析 王若遥 何晶晶 李盈洁[1] 彭婵娟 张晓[1] 冉黎 Niu Yingjie;Wu Panna;Hu Xi;Wang Ruoyao;He Jingjing;Li Yingjie;Peng Chanjuan;Zhang Xiao;Ran Li(Chengdu Medical College,Chengdu 610500,China)

机构地区：[1]成都医学院,成都610500

出　　处：《成都医学院学报》2018年第5期535-541,共7页Journal of Chengdu Medical College

基　　金：国家大学生创新创业训练项目(No:201613705037)

摘　　要：目的探讨内源性睫状细胞神经生长因子(ciliary neurotrophic factor,CNTF)和神经营养因子4(neurotrophin-4,NT-4)在大鼠脊髓损伤(spinal cord injury,SCI)早期的表达变化及其相互关系,为SCI神经营养因子疗法提供理论依据。方法成年Sprague Dawley(SD)大鼠45只随机分为假手术(Sham)组、SCI-12h、SCI-1d、SCI-3d和SCI-7d组(n=9)。各SCI组大鼠均采用改良Allen's法制备T10节段脊髓顿挫伤模型,用BBB(BassoBeattie-Bresnahan locomotor,BBB)评分法评估各组大鼠后肢运动功能;采用荧光定量PCR(Q-PCR)及免疫组织化学染色检测大鼠脊髓顿挫伤后CNTF与NT-4的表达情况;利用GeneMANIA数据库对CNTF与NT-4的相互作用进行预测分析。结果术后Sham组大鼠BBB评分不变,SCI组大鼠损伤平面以下完全瘫痪,BBB评分为0,SCI-7d组评分较SCI-12h组高(P<0.01),但仍低于Sham组(P<0.01)。与Sham组相比,SCI-12h组CNTF mRNA相对表达量降低(P<0.05),于SCI-3d组降至最低(P<0.05);NT-4mRNA相对表达量在SCI-12h组无变化,于SCI-1d组表达增高(P<0.05),至SCI-7d组表达降低(P<0.01),但与Sham组比较差异无统计学意义(P>0.05)。免疫组织化学染色结果显示,CNTF在神经元与神经胶质细胞均有表达,NT-4主要在神经元表达。与Sham组相比,CNTF与NT-4阳性神经元数量及IOD值均在SCI-12h组减少(P<0.01);两种蛋白质随后呈现不同的变化趋势:脊髓前角CNTF阳性神经元数量与IOD值分别在SCI-3d组与SCI-7d组再次降低(P<0.01);而NT-4阳性神经元与IOD值分别在SCI-7d组与SCI-3d组增高,表达水平接近Sham组。脊髓后角CNTF阳性神经元数量在SCI-1d组再次降低(P<0.01),至SCI-7d组无变化;NT-4阳性神经元数量及其IOD值分别在SCI-1d组及SCI-7d组增高(P<0.01),且SCI-7d组与Sham组比较差异无统计学意义(P>0.05)。GeneMANIA分析结果显示,CNTF和NT-4无直接相互作用,但与NGF、Tmem237、Kcnh6、Tph1、Psisp1和Ghrhr具有共同的共定位和/或共表达关系。结论机体在SCI急性期上�Objective To investigate the expression changes of endogenous ciliary neurotrophic factor （CNTF） and neurotrophin-4 （NT-4）in the spinal cord of the rat model with spinal cord injury （SCI） and analyze their correlation in order to provide the theoretical basis for the SCI neurotrophic factor therapy. Methods A total of 45 Sprague Dawley rats were randomly divided into the Sham-operated group and groups of SCI-12h, SCI-1d, SCI-3d and SCI-7d, and each group included 9 rats. After the T10 segmental SCI models were established with Allen＇s impact device in the SCI groups, the Basso-Beattie-Bresnahan （BBB） score was used to detect the motor function immediately, 12 hours, 1 day, 3 and 7 days respectively, and the fluorescence quantitative PCR and immunohistochemical staining were used to detect the expression changes of CNTF and NT-4 after spinal cord contusion, and the GeneMANIA database was used to predict the protein interaction of CNTF and NT-4. Results The BBB score of the SCI-7d group was significantly higher than that of the SCI-12h group （ P 〈0.01） and significantly lower than that of the sham-operated group （ P 〈0.01）. Compared with the sham-operated group, the relative expression level of CNTF mRNA decreased significantly in the SCI-12h group （ P 〈0.05） and reached the lowest in the SCI-3d group （ P 〈0.05）. The relative expression level of NT-4 mRNA didn＇t change in the SCI-12h group, while it increased significantly in the SCI-1d group （ P 〈0.05） and decrease significantly in the SCI-7d group （ P 〈0.05）. The relative expression level of NT-4 mRNA in the SCI groups were not significantly different from that of the sham-operated group. The results of the immunohistochemical staining showed that the CNTF positive cells were distributed in neurons and neuroglial cells and the NT-4 positive cells were mainly located in the cytoplasm of neurons. Compared with the sham-operated group, the number and integrated optical density （IOD） of CNTF and NT-4 posi

关 键 词：脊髓损伤 睫状神经营养因子 神经营养因子4 BBB评分 大鼠 

分 类 号：R651.2[医药卫生—外科学]