坐骨神经切断后胶质细胞源性神经营养因子mRNA的表达变化

目的探讨大鼠坐骨神经切断后,胶质细胞源性神经营养因子(GDNF)mRNA在两侧断端的表达变化及其意义。方法采用Trizol一步法提取坐骨神经总RNA,取少量作紫外扫描分析和琼脂糖电泳分析,切断SD大鼠左侧坐骨神经,不同时间、半定量RT-PCR方法,观察两侧断端GDNFmRNA的表达变化。结果坐骨神经切断前,GDNFmRNA在两侧坐骨神经微量表达,为(6.5±1.3)%,坐骨神经切断后远断端表达逐渐增加,伤后1d(7.8±1.9)%,伤后7d(10.3±2.1)%,伤后14d(11.9±2.3)%,伤后28d(12.3±2.4)%,近断端表达逐渐减少,伤后1d(5.8±1.3)%,伤后7d(4.0±1.0)%,伤后14d(3.6±1.1)%,伤后28d(3.1±1.0)%。伤后1d与伤前比较(P>0.05,t=1.193,0.879),伤后7,14,28d与伤前比较(P<0.01,t=3.762,3.565,5.059,4.083,5.164,4.905)。结论GDNF在损伤信号的刺激下表达急剧增加,起到修复神经元的作用,为外源性GDNF治疗脊髓损伤提供了理论基础。     

坐骨神经震荡伤后腰髓背根神经节病理改变及意义

目的　应用图像分析和免疫组织化学技术观察和分析兔坐骨神经高速弹丸震荡伤后腰髓背根神经节病理改变及意义。方法　大耳白兔 2 5只 (包括正常对照 5只 ) ,致伤靶点为右后肢外侧坐骨神经体表投影线中点 (0 38g钢珠 ,0 6 5g装药量 ) ,观察伤后 1、3、7、14d(n =5 )腰髓背根神经节病理改变及一氧化氮合酶 (nitricoxidesynthase ,NOS)表达变化 (免疫组化法 ) ,并进行神经元计数及神经元截面积图像分析。结果　腰髓背根神经节伤后发生出血、水肿、神经元皱缩、坏死等变化 ,伤后 3dNOS表达显著增强 ,伤后 7d神经元数显著减少 ,神经元平均截面积显著减少。结论　坐骨神经高速弹丸震荡伤后腰髓背根神经节发生了较重的损伤。     

大鼠脊髓背角TRPV4在背根神经节持续受压致神经病理性疼痛中的作用

目的:探讨大鼠背根神经节(dorsal root ganglion,DRG)持续受压(chronic compression of right side dorsal root ganglion,CCD)后脊髓背角瞬时感受器电位离子通道4(TRPV4)基因及蛋白变化,明确脊髓背角TRPV4在CCD致神经病理性疼痛中的作用。方法:采用健康成年雄性Wistar大鼠,共36只,随机分为3组,分别为空白对照组、CCD手术组、CCD+钌红组。制备大鼠背根神经节持续受压模型,于术前1天、术后第7天、给药前及给药2h后,测量大鼠机械刺激缩爪反应阈值,观察机械痛阈的变化;利用RT-PCR及Western Blot技术检测各组大鼠手术侧脊髓背角TRPV4基因及蛋白表达的变化。结果:与空白对照组相比,术后第7天,CCD组大鼠术侧机械痛阈值明显下降(P0.001),同侧脊髓背角TRPV4基因及蛋白表达升高(P0.05);与给药前相比,给予钌红2h后,术侧机械痛阈值明显升高(P0.001),同侧脊髓背角TRPV4基因及蛋白表达下降(P0.05)。结论:CCD后大鼠术侧机械痛阈下降,脊髓背角TRPV4基因及蛋白表达升高;钌红可部分逆转CCD后痛觉过敏,部分降低脊髓背角TRPV4基因及蛋白表达。脊髓背角TRPV4参与CCD后大鼠神经病理性疼痛形成。     

The morphological effects of a radio frequency lesion adjacent to the dorsal root ganglion (RF-DRG)--an experimental study in the goat.

Radiofrequency (RF) lesions adjacent to the dorsal root ganglion (DRG) are increasingly used in the treatment of intractable chronic pain of spinal origin. Opinions differ on which nerve fibres are affected by these lesions. Morphological studies have been carried out to assess the effects of radiofrequency lesions on nervous tissue. Interpretation has been difficult, since most studies have been performed under circumstances which are not comparable to the clinical situation. This study was undertaken to investigate morphological effects of RF lesions as they develop in the normal clinical situation. In two goats 22 G 100 mm SMK electrodes with a 5 mm active tip (Radionics) were positioned posterior to the lumber dorsal root ganglia (DRG). Sixty-second 67;C lesions were made on one side. The contralateral side was used as sham operation. The goats were sacrificed 2 weeks after the procedure. The lesions were studied for size as well as for effects on proliferation and regeneration using Ki-67 (MIB-1). Lesions made inside the DRG (23) were 1.8-2.0 mm in size. In these lesions there was a total loss of myelinated fibres. In lesions made adjacent to the DRG there was a significantly higher MIB-1 labelling on the treated side as compared to the sham-treated side. An RF lesion inside the DRG destroys myelinated fibres. A lesion adjacent to the DRG increases MIB-1 activity, indicating proliferation and regeneration after 2 weeks, despite the fact that the lesion was made outside the ganglion.     

低能量激光对大鼠背根神经节持续受压后痛觉敏感及TRPV4表达的影响

目的:观察低能量激光对大鼠背根神经节持续受压(chronic compression of the dorsal root ganglion,CCD)后痛觉敏感及脊髓背角内TRPV4表达及分布的影响.方法:选取健康雄性Wistar大鼠36只,分为空白组、CCD组、激光组各12只,各组均给予常规饲养.激光组在CCD后第4...     

直流电对鼠坐骨神经横断致L4-6脊髓前角运动神经元凋亡及bax基因表达的影响

目的 :观察直流电对鼠坐骨神经横断后脊髓运动神经元凋亡及bax表达的影响。方法 :将鼠右侧坐骨神经自梨状肌下缘 0 .5cm处切断 ,实验组在相应的脊髓节段立即置入直流电刺激器 ,对照组置入无输出电流的电刺激器 ,手术后 1、4、7、14、2 8d处死动物 ,切取L4— 6段脊髓 ,对右侧脊髓阳性运动神经元计数 ,应用免疫组化技术 ,对右侧脊髓bax表达阳性运动神经元计数。结果 :坐骨神经切断后 4、7、14d ,实验组脊髓Tunel染色阳性运动神经元数明显少于对照组 ,术后 4、7、14、2 8d ,实验组脊髓bax阳性运动神经元数明显少于对照组。结论 :直流电刺激对坐骨神经切断引起的脊髓运动神经元凋亡的预防作用与直流电刺激抑制神经元的bax表达有关。     

Upregulation and redistribution of ephrinB and EphB receptor in dorsal root ganglion and spinal dorsal horn neurons after peripheral nerve injury and dorsal rhizotomy

EphrinB–EphB receptor signaling plays diverse roles during development, but recently has been implicated in synaptic plasticity in the matured nervous system and in pain processes. The present study investigated the correlation between expression of ephrinB and EphB receptor proteins and chronic constriction injury (CCI) of the sciatic nerve and dorsal rhizotomy (DR) in dorsal root ganglion (DRG) and spinal cord (SC); and interaction of CCI and DR on expression of these signals. Adult, male Sprague–Dawley rats were employed and thermal sensitivity was determined in the sham operated CCI and DR rats. Western blot and immunobiochemistry analysis and immunofluorescence staining techniques were used to detect the expression and location of the ephrinB–EphB receptor proteins in DRG and SC. The results showed that expression of ephrinB1 and EphB1 receptor proteins was significantly upregulated in DRG and SC in a time‐dependent manner corresponding to the development of thermal hyperalgesia after CCI. The increased expression is predominately located in the medium‐ and small‐sized DRG neurons, the superficial layers of spinal dorsal horn (DH) neurons, and the IB4 positive nociceptive terminals. DR increases ephrinB1 in DRG, not SC and EphB receptor in SC, not DRG. DR suppressed CCI‐induced upregulation of ephrinB1 in SC and EphB1 receptor in DRG and SC. These findings indicate that ephrinB–EphB receptor activation and redistribution in DRG and DH neurons after nerve injury could contribute to neuropathic pain. This study may also provide a new mechanism underlying DR‐induced analgesia in clinic.     

液压冲击脑损伤大鼠海马BDNF mRNA表达实验研究

目的：研究脑损伤后脑脑源性神经营养因子（BDNF）mRNA表达及其时序性变化,进一步研究脑损伤的分子机制。方法：用免疫组织化学及原位杂交方法,观察液压冲击脑损伤大鼠海马BDNF mRNA受伤后不同时间（5、15、30分钟和1、3、6、12小时及1、3、7日）的表达情况,以正常大鼠及手术大鼠作为对照。结果：正常及手术对照组大鼠脑组织海马表达少量的BDNF;脑损伤后1小时海马齿状回CA2及CA3细胞表达增强,3－6小时达高峰,12小时开始下降,24小时后降至对照水平。1、3、6和12小时与正常及手术对照组比较均有显著性差异。结论：液压冲击脑损伤可诱导BDNF基因的表达,BDNF mRNA表达存在时间差异,提示机体对脑损伤存在自身保护机制。     

甲泼尼龙联合注射用鼠神经生长因子治疗急性脊髓损伤的临床应用及护理

目的研究和观察急性脊髓损伤（ASCI）患者应用甲泼尼龙联合注射用鼠神经生长因子（鼠NGF）的治疗方案的价值及护理。方法选取急性脊髓损伤患者共53例,分为治疗组33例和对照组20例,治疗组术后采用甲泼尼龙联合注射用鼠NGF的治疗方案,对照组为未采用此方案的常规治疗方法。对2组患者均实施整体护理方案,按日本骨科协会（JOA）评分计算患者术前、术后1周、术后1年的神经功能恢复率,并统计并发症发生情况,远期神经功能评分增加按ODOM分级评分。结果2组患者组间术前JOA评分无统计学差异（P〉O．05）;2组1年后随访未发现并发症,植骨融合良好,内固定稳定;治疗组术后5d发生感染1例,经抗炎抗生素治疗后治愈。与术前相比,2组术后1周及1年JOA评分均显著增加,治疗组改善率显著高于对照组,差异有统计学意义（P〈0．01）;1年后治疗组ODOM评分优良率显著高于对照组,差异有统计学意义（P〈0．01）。结论甲泼尼龙联合注射用鼠NGF治疗急性脊髓损伤的同时配合以优质的护理,可改善急性脊髓损伤患者术后的疼痛,促进神经功能的恢复,对于改善患者生活质量具有显著意义。