神经丝氨酸蛋白酶抑制剂(NSP)对大鼠急性脊髓损伤(SCI)后神经功能的修复作用  

Repair effects of neuroserpin (NSP) on neural function after acute spinal cord injury (SCI) in rats

作　　者：吕越昌 施德源[1] 姜晓幸[1] 冯振洲[1] 蒋淳[1] 陈子贤[1] 

机构地区：[1]复旦大学附属中山医院骨科,上海200032

出　　处：《复旦学报（医学版）》2014年第2期216-221,共6页Fudan University Journal of Medical Sciences

摘　　要：目的通过检测脑源性神经生长因子(brain-derived neurotrophic factor,BDNF)、胶质细胞源性神经生长因子(glial cell line-derived neurotrophic factor,GDNF)和轴突生长抑制因子(neurite outgrowth inhibitor A,Nogo-A)的表达情况,来评估神经丝氨酸蛋白酶抑制剂(neuroserpin,NSP)对急性脊髓损伤(spinal cord injury,SCI)大鼠神经功能的恢复情况并对其机制进行初步探索。方法 130只成年雄性SD大鼠随机分成3组:SCI组(n=60),用改良Allen′s重物打击法制备SCI模型,鞘内注射生理盐水25μL。NSP组(n=60),大鼠SCI后立即鞘内注射NSP 25μL。正常对照组(n=10),只去除椎板,暴露脊髓,不造成SCI。各组定期行为学观察(BBB评分),Western blot检测SCI局部GDNF和Nogo-A的表达,real-time PCR检测SCI局部BDNF、GDNF和NogoA的表达情况。结果随时间延长,术后SCI组和NSP组大鼠BBB评分均逐渐升高,但是两组并无明显差异。Western blot和real-time PCR显示:急性SCI后1天GDNF表达明显升高,且随时间延长其表达量逐渐降低,同时,在1、3、7和14天各时间点NSP组GDNF表达量均高于SCI组。急性SCI后Nogo-A表达成逐渐升高趋势,同时,NSP组在各时间点的Nogo-A表达量均低于SCI组。real-time PCR显示:SCI 1天后BDNF表达水平显著提高,随时间推移,表达水平逐渐下降;同时,NSP组在1、3、7和14天各时间点BDNF水平均明显低于SCI组。结论急性SCI后,大鼠内源性BDNF、GDNF和Nogo-A的表达水平均升高。外源性给予NSP对大鼠急性SCI后的运动功能无明显改善,但在基因和蛋白质水平对大鼠的神经功能恢复有一定的促进作用。Objective To explore the expressions of brain-derived neurotrophic factor （BDNF）,glial cell linederived neurotrophic factor （GDNF） and neurite outgrowth inhibitor A （Nogo-A） in rat spinal cord tissues with acute spinal cord injury （SCI）,and then to study whether neuroserpin（NSP） could change these expressions to protect the neural function.Methods One hundred and thirty SD rats were randomly divided into 3 groups：SCI group （n=60）,NSP group （n=60） and control group （n=10）.The acute SCI models were made according to the Allen′s weight drop method.SCI group were injected with 25 μL normal saline,and NSP group were treated with 25 μL NSP by intrathecal injection immediately after acute SCI.In control group,we only removed the vertebral plate and did not injure the spinal cord.Then we used Basso Beattie Bresnahan（BBB） scores to evaluate the functional deficits of the spinal cord.Western blot and realtime PCR were used to detect the expressions of BDNF,GDNF and Nogo-A.Results There were no significant difference in BBB scores between SCI and NSP group.Western blot and realtime PCR both indicated that GDNF expressions increased significantly one day after acute SCI compared with control group,and declined thereafter.GDNF expressions of NSP group were higher than these of SCI group at 1,3,7 and 14 d respectively after acute SCI.Nogo-A expressions after acute SCI increased compared with control group,and NSP group was lower than SCI group in every time point respectively.Meanwhile,real-time PCR showed that BDNF expressions after acute SCI reached the highest at 1 day and declined thereafter.However,the BDNF expressions of NSP group were significantly lower than these of SCI group at 1,3,7 and 14 d after acute SCI respectively.Conclusions The expressions of BDNF,GDNF and Nogo-A after ASCI all increased compared with control group.There were no significant improvement in the motor function of ASCI rats with NSP by intrathecal injection,but NSP could promote the neural

关 键 词：神经丝氨酸蛋白酶抑制剂(NSP) 脊髓损伤(SCI) 脑源性神经生长因子(BDNF) 胶质细胞源性神经生长因子(GDNF) 轴突生长抑制因子(Nogo—A) 大鼠 

分 类 号：R683.2[医药卫生—骨科学]