抑制脊髓络丝蛋白表达对神经性疼痛成年大鼠疼痛行为的影响研究  

Effects of the Inhibition of Reelin Expression in Spinal Cord on Pain Behaviors in Neuropathic Pain Adult Rat Models

作　　者：胡洁 旷昕[1] 张倩璐[1] 李欢颜[1] 阳巧云 彭良玉[1] HU Jie;KUANG Xin;ZHANG Qianlu;LI Huanyan;YANG Qiaoyun;PENG Liangyu(Department of Anesthesiology,the First Affiliated Hospital of University of South China,Hengyang 421001,China)

机构地区：[1]南华大学附属第一医院麻醉科

出　　处：《中国全科医学》2019年第20期2448-2454,共7页Chinese General Practice

基　　金：国家自然科学基金青年项目(81300971);湖南省卫生计生委科研计划项目(B20180203)

摘　　要：背景络丝蛋白是主要参与神经元发育和成熟大脑突触过程的大分子细胞外基质糖蛋白。在神经元迁移过程终止后,脊髓尤其是脊髓背角浅层的神经元中仍有络丝蛋白的表达。神经性疼痛大鼠脊髓背角中络丝蛋白的表达明显减少,但其在伤害性感受和疼痛中的作用仍不明确。目的探讨鞘内注射络丝蛋白shRNA对成年大鼠疼痛行为的影响。方法 2015年,选取8~10周龄的SPF级雄性SD大鼠119只,置管成功108只,采用随机分组法将其分为正常(Norm)组12只、RNA干扰(RNAi)组12只、假手术(Sham)组12只、Sham+RNAi(SR)组6只、慢性坐骨神经压迫性损伤(CCI)组27只、CCI+RNAi(CR)组27只、CCI+空载体(EV)组12只。CCI组、CR组和EV组制备CCI模型,RNAi组、SR组、CR组鞘内给予络丝蛋白shRNA慢病毒载体,EV组给予空载体。计算干预后第4、7、10、14、21天络丝蛋白表达水平,于干预后第10天进行免疫组织化学观察络丝蛋白定位情况,检测干预前及干预后第1、4、7、10、14、17、21天大鼠足底机械缩足阈值(PWMT)、热辐射刺激潜伏期(PWTL)。结果 RNAi组第10天左、右侧络丝蛋白表达水平均高于Norm组,CR组第10天左侧络丝蛋白表达水平低于Sham组、CCI组,CCI组、CR组、EV组第10天右侧络丝蛋白表达水平均低于Sham组,CR组第10天右侧络丝蛋白表达水平低于CCI组,CCI组第4天右侧络丝蛋白表达水平低于CCI组左侧,CR组第4天右侧络丝蛋白表达水平低于CR组左侧,CCI组右侧、CR组左侧第7、10、14、21天络丝蛋白表达水平均低于CCI组左侧,CR组右侧第7、10、14、21天络丝蛋白表达水平均低于CCI组右侧(P<0.05)。Norm组络丝蛋白弥漫性分布于脊髓背角Ⅰ~Ⅱ、Ⅴ板层和外侧脊髓核(LSN);RNAi组双侧脊髓背角的Ⅰ~Ⅱ板层及LSN中仅有极少量络丝蛋白,Ⅴ板层仍存少量表达;Sham组双侧脊髓背角中络丝蛋白表达则无明显影响;CCI组右侧脊髓背角中络丝蛋白表达水平低�Background Reelin is a large secreted extracellular matrix glycoprotein that modulates neuronal development and participates to the functioning of adult central synapses.After migrations have ceased,neurons within the spinal cord especially in the superficial dorsal horn continue to express Reelin.The expression of spinal dorsal horn Reelin downregulates significantly in neuropathic pain rats,while the role of Reelin in nociceptive processing and pain is still unknown.Objective To investigate the effects of intrathecal injection of Reelin shRNA on pain behaviors in adult rats.Methods A total of 119 8-10-week-old healthy male SD rats of SPF grade were selected and treated with intrathecal catheter implantation in 2015.The 108 rats with successful implantation were randomly assigned to normal group(Norm gourp,n=12),RNA interference group(RNAi group,n=12),sham-operation group(Sham group,n=12),sham plus RNA interference group(SR group,n=6),chronic compression injury(CCI) of sciatic nerve group(CCI group,n=27),CCI plus RNA interference group(CR group,n=27),and CCI plus empty vector group(EV group,n=12),respectively.Groups of CCI,CR and EV received a partial sciatic nerve ligation to establish CCI models.RNAi,SR and CR groups were intrathecally treated with lentiviral vector of Reelin shRNA,while EV group received intrathecal injection of an empty lentiviral vector.Western blotting was used to test the expression level of Reelin protein on the 4 th,7 th,10 th,14 th and 21 st days of intervention.Immunohistochemical technique was used to observe the localization of Reelin in the spinal dorsal horn on the 10 th day of intervention.Paw withdrawal mechanical threshold(PWMT) and paw withdrawal thermal latency(PWTL) were tested before and on the 1 st,4 th,7 th,10 th,14 th,17 th and 21 st days of intervention,respectively.Results On the 10 th day of intervention,compared with rats in Norm group,the mean expression level of Reelin protein increased significantly on both right and left sides of spinal dorsal horn of rats in RNA

关 键 词：神经痛 疼痛 络丝蛋白 RNA干扰 痛觉过敏 

分 类 号：R745[医药卫生—神经病学与精神病学]