神经生长因子对创伤性脊髓损伤大鼠膀胱功能和轴突损伤修复的作用研究  

The improvement of bladder function and axon repair in rats with traumatic spinal cord injury via MAPK/ERK pathway by nerve growth factor

作　　者：袁绍港 谢晶军[2] 范彦瑞 王朝亮[3] Yuan Shaogang;Xie Jingjun;Fan Yanrui(Department of Rehabilitation Medicine,the Second Affiliated Hospital of Henan University of Science and Technology,Luoyang Henan 471000;不详)

机构地区：[1]河南省洛阳市河南科技大学第二附属医院康复医学科,471000 [2]湖州市第一人民医院康复医学科 [3]郑州大学第一附属医院泌尿外科

出　　处：《卒中与神经疾病》2022年第3期230-235,共6页Stroke and Nervous Diseases

基　　金：2019年度河南省医学科技攻关计划联合共建项目(LHGJ20190175)。

摘　　要：目的 探讨神经生长因子(Nerve growth factor,NGF)对创伤性脊髓损伤(Traumatic spinal cord injury,t-SCI)大鼠膀胱功能和脊髓神经轴突损伤修复的影响及其分子机制。方法 取30只雄性Sprague-Dawley(SD)大鼠,通过改良Allen’s击打法构建创伤性脊髓损伤模型,随机分为假手术组、损伤组和NGF组,每组各10只;采用血脑屏障(Blood-brain barrier,BBB)评分观察术前、术后大鼠的后肢运动功能;BL-420生物仪实验系统检测尿动力学;甲苯胺蓝染色吻合口远端截取的左侧腰6前根,计算有髓轴突数量;采用苏木精-伊红(Hematoxylin eosin,HE)染色大鼠膀胱组织;原位末端标记法(TdT-mediated dUTP nick and labeling,TUNEL)染色大鼠损伤严重的脊髓,观察脊髓神经细胞的凋亡率;蛋白免疫印迹法(Western blot)检测脊髓组织中原癌基因丝氨酸/苏氨酸蛋白激酶(proto-oncogene serine/threonine-protein kinase,Raf-1)、磷酸化丝裂原活化蛋白激酶激酶(Phospho-mitogen-activated protein kinase kinase 2,p-MEK-2)、磷酸化丝裂原活化蛋白激酶激酶(Mitogen-activated protein kinase kinase 2,MEK-2)、磷酸化细胞外调节蛋白激酶(Phospho-extracellular regulated protein kinases1/2,p-ERK1/2)、细胞外调节蛋白激酶(Extracellular regulated protein kinases1/2,ERK1/2)的蛋白表达水平。结果 BBB评分显示,术前假手术组、损伤组和NGF组的评分无明显差异,而术后与假手术组比较,损伤组和NGF组的评分显著降低(P<0.05);损伤组的最大逼尿压力和脊髓有髓轴突数量显著低于假手术组和NGF组(P<0.05),但损伤组的残余尿量和膀胱细胞凋亡率显著高于假手术组和NGF组(P<0.05);HE染色显示,损伤组的膀胱水肿严重,逼尿肌结构疏松,而NGF组膀胱损伤减轻;Western blot显示,与假手术组比较,损伤组的p-ERK1/2/ERK1/2和p-MEK-2/MEK-2蛋白水平比值以及Raf-1表达水平显著高于假手术组和NGF组(P<0.05);假手术组和NGF组最大逼尿压力、髓轴突数量、残余尿�Objective To explore the effects of nerve growth factor(NGF) on bladder function and axon repair in rats with traumatic spinal cord injury(t-SCI),and explore its underlying molecular mechanism.Methods Thirty male SD rats were established the traumatic spinal cord injury model by modified Allen’s beating method.They were randomly divided into sham operation group,injury group and NGF group,with 10 rats in each group.We used the BBB score to observe the motor function of the hind limbs of the rats before and after the operation.The BL-420 biometer experimental system was used to detect the urodynamics.The sixth anterior root of the left lumbar nerves taken from the distal end of the anastomosis was stained with toluidine blue,and the number of myelinated axons was counted.HE was used to stain rat bladder tissues.The severely injured spinal cords were stained with TUNEL and the apoptosis rate was evaluated.Western blot was applied to detect proto-oncogene serine/threonine-protein kinase(Raf-1),phospho-mitogen-activated protein kinase kinase 2(p-MEK-2),mitogen-activated protein kinase kinase 2(MEK-2),phospho-extracellular regulated protein kinases1/2(p-ERK1/2) and extracellular regulated protein kinases1/2(ERK1/2) expression in spinal cord tissues.Results The BBB score results showed that there was no difference in the scores of the sham operation group,the injury group and the NGF group before the operation.After the operation,the scores of the injury group and the NGF group were significantly lower than the sham operation group(P0.05).Conclusion NGF may hinder the conduction of MAPK/ERK pathway,thereby affecting the repair of injured axons and improving the bladder function of t-SCI rats.

关 键 词：创伤性脊髓损伤 神经生长因子 膀胱 轴突 大鼠 

分 类 号：R651.2[医药卫生—外科学]