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

研究设计。使用核因子-kB诱饵的大鼠炎性疼痛模型的体外和体内研究。摘要和背景数据。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。我们假设用核因子-kB诱饵抑制核因子-kB基因的表达可能会抑制炎性疼痛。在组织块培养中诱导NF-kB诱骗FITC，将其注入坐骨神经内，检测其对DRG的转导效率。行为学测试：12只大鼠足底注射完全弗氏佐剂，随机分为3组：诱饵组-单次神经内注射10111的核因子-kB诱饵(n=4)，生理盐水组-单次神经内注射10μL生理盐水(n=4)，单纯组-不治疗(n=4)。使用von Frey灯丝和带有热源的Hargreaves装置进行行为测试。体外转导效率为53.6%，体内转导效率为20.5%。所有FITC阳性神经元的细胞大小分布类型没有统计学差异。在行为学测试中，在机械性痛觉过敏实验中，假手术组与生理盐水组在术后2~14d，在热痛觉过敏实验中，在术后2~3d，假手术组与生理盐水组的戒断潜伏期或阈值有显著差异。在体内和体外将核因子-kB诱骗基因转导入背根神经节。此外，核因子-kB诱骗可减少炎性痛模型大鼠的机械性痛觉过敏和热痛觉过敏，提示核因子-kB诱骗抑制核因子-kB可能是介导炎症或减轻炎性疼痛的关键机制。

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.