Study of the gene therapy with extracorporeal shockwaves for Discogenic low back pain

研究设计。核因子- κ B （NF-kB）诱饵诱导大鼠炎性疼痛模型的体内外研究。摘要和背景资料。据报道，转录因子NF-kB在调节促炎细胞因子基因表达中起着至关重要的作用。我们假设用NF-kB诱饵抑制NF-kB基因表达可能抑制炎症性疼痛。通过外植体培养诱导NF-kB诱饵fitc，经神经内注射至坐骨神经，测定其向DRG的转导效率。行为学测试，12只大鼠接受足底完全弗氏佐剂注射，分为3组：诱饵组-单次神经内注射10111 NF-kB诱饵（n=4）；生理盐水组：单次神经内注射生理盐水10μl (n=4)；未治疗组（n=4）。使用von Frey灯丝和带热源的Hargreaves装置进行行为测试。NF-kB诱饵- fitc在体外和体内的总转导效率分别为53.6%和20.5%。所有fitc阳性神经元的细胞大小分布类型无统计学差异。在行为学测试中，假药组和生理盐水组在术后2 ~ 14天的机械异常性疼痛实验和术后2 ~ 3天的热痛觉过敏实验中，戒断潜伏期或阈值差异有统计学意义。在体内和体外均将NF-kB诱饵传递并转导到DRG中。此外，NF-kB诱骗物减少了大鼠炎症性疼痛模型中的机械性异常痛和热痛觉过敏，这表明NF-kB诱骗物抑制NF-kB可能是介导炎症或减轻炎症性疼痛的关键机制。

Study Design. In vitro and in vivo study of a rat inflammatory pain model using nuclear factor-kappa B (NF-kB) decoy.Summary and Background Data. Transcription factor NF-kB is reported to play a crucial role in regulating proinflammatory cytokine gene expression. We hypothesized that inhibiting NF-kB gene expression with NF-kB decoy may suppress inflammatory pain.Methods. NF-kB decoy-FITC was induced in explant culture, endoneurally injected into the sciatic nerve, and its transduction efficiency into DRG measured. For behavioral testing, 12 rats received plantar injections of complete Freund's adjuvant and were divided into three groups : decoy group - single endoneural injection of 10111 of NF-kB decoy (n=4) ; saline group - single endoneural injection of 10μl of saline (n=4) ; and naive group - untreated (n=4). Behavioral testing was performed using von Frey filaments and a Hargreaves device with a heat source.Results. Total transduction efficiency of NF-kB decoy-FITC was 53.6% in vitro and 20.5% in vivo. No statistical differences were observed with respect to types of cell size distributions of all FITC-positive neurons. In behavioral testing, withdrawal latencies or thresholds significantly differed between the decoy group and the saline group from 2 to 14 days after surgery in the mechanical allodynia experiments, and from 2 to 3 days after surgery in the thermal hyperalgesia experiments.Conclusions. NF-kB decoy was conveyed and transduced into DRG both in vivo and in vitro. Additionally, NF-kB decoy reduced mechanical allodynia and thermal hyperalgesia in the rat inflammatory pain model, suggesting that inhibition of NF-kB with NF-kB decoy may represent a key mechanism for mediating inflammation or reducing inflammatory pain.