CAJ | 학술논문

目的观察大鼠脊髓背角中NF-κB及其下游肿瘤坏死因子α(TNF-α)在慢性坐骨神经挤压性损伤(CCI)疼痛模型中表达的变化规律.方法雄性SD大鼠76只,随机分为手术组(CCI组)和假手术组(Sham组)(n=38),于CCI前1 d、CCI后1、4、7、14、21、28 d各时间点测定机械痛阈及热痛阈后立即处死大鼠,取腰髓,采用实时荧光定量PCR和免疫荧光双标的方法分别测定NF-κB和TNF-α的mRNA含情和蛋白表达变化.结果 CCI组大鼠手术侧机械痛阈及热痛阈明显降低,脊髓背角中NF-κB水平和TNF-α的表达增加,明显高于对侧和假手术组;NF-κB和TNF-α的mRNA水平于CCI后4 d开始增加,分别为术前的(1.20±0.16,2.32±0.27)倍,7 d达到高峰,为术前的(1.75±0.20,3.38±0.41)倍,随后TNF-α迅速下降,而NF-κB于CCI后28 d仍为术前的(1.15±0.24)倍,维持于较高水平(P＜0.05).术后第7天的免疫荧光双标显示NF-κB和TNF-α在术侧脊髓后角存在共定位表达.结论 CCI致大鼠脊髓背角NF-κB活化并上调TNF-α的表达,参与神经病理性疼痛的调控过程.
목적 관찰대서척수배각중NF-κB급기하유종류배사인자α(TNF-α)재만성좌골신경제압성손상(CCI)동통모형중표체적변화규률.방법 웅성SD대서76지,수궤분위수술조(CCI조)화가수술조(Sham조)(n=38),우CCI전1 d、CCI후1、4、7、14、21、28 d각시간점측정궤계통역급열통역후립즉처사대서,취요수,채용실시형광정량PCR화면역형광쌍표적방법분별측정NF-κB화TNF-α적mRNA함정화단백표체변화.결과 CCI조대서수술측궤계통역급열통역명현강저,척수배각중NF-κB수평화TNF-α적표체증가,명현고우대측화가수술조;NF-κB화TNF-α적mRNA수평우CCI후4 d개시증가,분별위술전적(1.20±0.16,2.32±0.27)배,7 d체도고봉,위술전적(1.75±0.20,3.38±0.41)배,수후TNF-α신속하강,이NF-κB우CCI후28 d잉위술전적(1.15±0.24)배,유지우교고수평(P＜0.05).술후제7천적면역형광쌍표현시NF-κB화TNF-α재술측척수후각존재공정위표체.결론 CCI치대서척수배각NF-κB활화병상조TNF-α적표체,삼여신경병이성동통적조공과정.
Objective To investigate the expressions of NF-κB and TNF-α in lumbar spinal cord in a rat model of chronic constrictive injury(CCI).Methods Seventy-six male SD rats were randomly divided into 2 groups(n=38 each):CCI group receiving chronic constriction injury and sham group receiving sham operation as control.The mechanical and thermal nociceptive thresholds were assessed with paw withdrawal latency(PWL)to von Frey filaments and radiant heat at difierent time points.Five animals were sacrificed at each time point for real-time polymerase chain reaction(real-time PCR)and another three animals sacrificed at 7 d post-operation for double-immunofluorescence histochemical staining.Lumbar segments of spinal cord were removed.The expressions of NF-κB and TNF-α in spinal cord were examined by real-time PCR and double-immunofluorescence histochemical technique.Results The post-operative thresholds to mechanical and thermal stimuli decreased obviously.As compared with contralateral side and sham group,the expressions of NF-κB and TNF-α mRNA increased significantly in ipsilateral spinal dorsal horn.Their expressions began to increase at 4 d post-operation and peaked at 7 d.Then TNF-α began to decrease while NF-κB maintained at a high level throughout the experiment.Double-immunofluorescence histochemical staining revealed extensive co-localization of NF-κB with TNF-α on ipsilateral side of dorsal horn.Conclusion The activation of NF-κB and its downstram inflammatory mediators may be involved in the regulation of neuropathic pain.