中文摘要：

目的 探讨Toll样受体4（TLR4）/NF-κB信号转导通路在大鼠骨癌痛形成及维持中的作用.方法 健康雌性SD大鼠81只,5～6周龄,体重150～ 180 g,采用随机数字表法,将其分为8组,骨癌痛组（BCP组）18只,其他7组每组9只,对照组（C组）不作任何处理；骨癌痛组大鼠胫骨注射l×105 Walker 256乳腺癌细胞；Ⅴ1组、TLR4siRNA1组和TLR4msiRNA1组分别于造模后4d（预注射）鞘内注射溶媒、TLR4小干扰RNA （TLR4siRNA）和TLR4错义siRNA（TLR4msiRNA）,Ⅴ2组、TLR4siRNA2组和TL-R4msiRNA2组分别于造模后9d（后注射）鞘内注射溶媒、TLR4siRNA和TLR4msiRNA.TLR4siRNA和TLR4msiRNA 2μg用5％葡萄糖和jetPEITM稀释到5μl,1次/d,连续3d,最后1次注射后24 h处死大鼠,BCP组分别于造模后7、12d处死大鼠,取L4-6脊髓,采用RT-PCR法检测NF-κBp65 mRNA、IL-6 mRNA和TNF-α mRNA表达,采用免疫组化法和Western blot法检测NF-κBp65表达.结果 预注射TLR4siRNA可完全抑制骨癌痛大鼠脊髓组织NF-κBp65 mRNA、TNF-α mRNA和NF-κBp65表达上调,而后注射TLR-4siRNA可部分抑制骨癌痛大鼠脊髓组织NF-κBp65 mRNA、IL-6 mRNA、TNF-α mRNA和NF-κB表达上调（P＜ 0.05或0.01）.结论 TLR4/NF-κB信号转导通路介导了大鼠骨癌痛的形成和维持.

英文摘要：

Objective To evaluate the role of Toll-like receptor 4 （TLR4）/NF-κB signal transduction pathway in development and maintenance of bone cancer pain （BCP） in rats.Methods Eighty-one female Sprague-Dawley rats,aged 5-6 weeks,weighing 150-180 g,were randomly divided into 8 groups：control group （group C）,BCP group,vehicle 1 group （Ⅴ1 group）,TLR4 small interfering RNA （siRNA） 1 group （TLR4siRNA1 group）,TLR4 mismatch siRNA （msiRNA） 1 group （TLR4msiRNA1 group）,Ⅴ2 group,TLR4siRNA2 group and TLR4msiRNA2 group.There were 18 rats in group BCP and 9 rats in each group of the left 7 groups.BCP was induced by intra-tibial inoculation of 1 × 105 Walker 256 breast cancer cells.In Ⅴ1,TLR4siRNA1 and TL-R4msiRNA1 groups,vehicle,TLR4siRNA and TLR4msiRNA were intrathecally injected,respectively,on 4th day after inoculation （pre-injection）.In Ⅴ2,TLR4siRNA2 and TLR4msiRNA2 groups,vehicle,TLR4siRNA and TLR4msiRNA were intrathecally injected,respectively,on 9th day after inoculation （post-injection）.TLR4siRNA and TLR4msiRNA 2 μg were diluted with 5% glucose and jetPEITM to 5 μl,and then given once a day for 3 consecutive days,and the rats were sacrificed at 24 h after the last injection.The rats in group BCP were sacrificed ondays 7 and 12 after inoculation.The lumbar segments （L4-6） of the spinal cord were removed for determination of the expression of NF-κBp65 mRNA,IL-6 mRNA and TNF-α mRNA （by real-time PCR） and NF-κBp65 （by Western blot and immuno-histochemistry）.Results Pre-injection of TLR4siRNA could completely inhibit up-regulation of the expression of NF-κBp65 mRNA,NF-κBp65 and TNF-α mRNA in the spinal cord tissues,however,post-injection of TLR4siRNA could partially inhibit up-regulation of the expression of NF-κBp65 mRNA,NF-κBp65,IL-6 mRNA and TNF-α mRNA in the spinal cord tissues （P 〈 0.05 or 0.01）.Conclusion TLR4/NF-κB signaling pathway mediates the development and maintenance of BCP in rats.