脊髓NRSF在小鼠骨癌痛形成中的作用及其与NR2B表达的关系  

Role of spinal cord neuronal restrictive silencing factor in development of bone cancer pain and its relationship with N-methyl-D-aspartate receptor subunit 2B expression in mice

作　　者：王丹 余剑波 

机构地区：[1]天津市中西医结合医院南开医院麻醉科,300100

出　　处：《中华神经医学杂志》2017年第6期576-580,共5页Chinese Journal of Neuromedicine

摘　　要：目的探讨脊髓神经元限制性沉默因子（NRSF）在小鼠骨癌痛形成中的作用及其与N-甲基-D-天冬氨酸（NMDA）受体2B亚基（NR2B）表达的关系。方法SPF级成年雄性C3H／HeJ小鼠96只采用随机数字表法分为4组：骨癌痛组（n=241、NRSF反义脱氧寡核苷酸组（AS-ODN组，n=24）、人工脑脊液组（n=24）和假手术组（n=24）。前3组将含2×10^5个NCTC2472纤维肉瘤细胞的混悬液（含最低有效的α-MEM液体培养基）接种至小鼠右侧股骨远端骨髓腔制备骨癌痛模型，假手术组接种不含NCTC2472纤维肉瘤细胞的α-MEM液体培养基。AS—ODN组于接种前及接种后2d、4d和6d鞘内注射含AS-ODN10μg的人工脑脊液5μL，人工脑脊液组注射等体积人μ脑脊液。（1）各组小鼠于接种前（T0）及接种后5d（T1）、7d（T2）、10d（T3）和14d（T4）时分别测定机械缩足阈值（PWMT）和热刺激缩足反应潜伏期（PWTL）。（2）各组小鼠于T1、T2、T3和T4时间点分别处死取脊髓腰膨大，采用Westernblotting检测NRSF及NR2B蛋白表达。结果（1）与假手术组比较，骨癌痛组及人工脑脊液组T2、T3、T4时间点PWMT明显降低，T3、T4时间点PWTL明显降低，差异均有统计学意义（P〈0．05）；AS—ODN组T1、T2、T3、T4时间点PWMT及PWTL均明冠降低，差异均有统计学意义（P〈0．05）。与骨癌痛组比较，AS-0DN组T1、T2时间点PWMT及PWTL均明显降低，差异均有统计学意义（P〈0．05）。（2）与假手术组比较，骨癌痛组、AS—ODN组及人工脑脊液组T1、T2、T3、T4时间点NR2B蛋白表达明显增高，NRSF蛋白表达明显降低，差异均有统计学意义（P〈0．05）。与骨癌痛组比较，AS-0DN组T1、T2时间点NR2B蛋白表达明显增高，NRSF蛋白表达明姓降低，差异均有统计学意义（P〈0．05）。结论脊髓NRSF在小鼠骨癌痛形成中具有重要作用，其对NR2B的渊控作用参与了骨癌痛的形成。Objective To evaluate the effect of spinal cord neuronal restrictive silencing factor （NRSF） in the development of bone cancer pain and its relationship with N-methyl-D-aspartate receptor subunit 2B （NR2B） in mice. Methods Ninety-six SPF adult male mice （C3H/HeJ） were randomly divided into bone cancer pain group （Group BCP, n=24）, NRSF antisense oligodeoxynucleotide （AS-ODN） group （Group AS-ODN, n=24）, artificial cerebrospinal fluid （ASCF） group （Group ACSF, n=24） and sham-operated group （Group S, n=24）. Suspension with 2×10^5 NCTC2472 fibrosarcoma cells were injected into the medullary cavity of right distal femur to induce bone cancer pain of mouse models in the former three groups; mice in Group S were injected only a-MEM; before and 2, 4 and 6 d after inoculation, mice in the Group AS-ODN were given 5 μL artificial cerebrospinal fluid containing AS-ODN 10 μg via intrathecal injection, and mice in the Group ACSF were given ACSF only. （1） All mice were tested for pain-related behaviors before inoculation （TO） and 5 d （T1）, 7 d （T2）, 10 d （T3） and14 d （T4） after inoculation, including paw withdrawal mechanical threshold （PWMT） and paw withdrawal thermal latency （PWTL）. （2） Lumbar intumescentia of mice in all groups was taken to investigate the expression levels of NR2B and NRSF by Western blotting after pain behavior tests at T1, T2, T3 and T4 time points. Results （1） As compared with that in Group S, the PWMT in Group BCP and ACSF was significantly decreased at T2, T3 and T4 time points, while PWTL was significantly decreased at T3 and T4 time points （P〈0.05）; both PWMT and PWTL in Group AS-ODN were significantly decreased as compared with those in Group S at T1, T2, T3 and T4 （P〈0.05）; as compared with those in the Group BCP, PWMT and PWTL in Group AS-ODN decreased significantly at T1 and T2 time points （P〈0.05）. （2） As compared with Group S, the NR2B expression in Group BCP, AS-ODN and ACSF was significantly i

关 键 词：骨肿瘤 慢性疼痛 神经元限制性沉默因子 N-甲基-D-天冬氨酸受体2B观基 

分 类 号：R738.1[医药卫生—肿瘤]