腰椎间盘突出症(LDH)是一种常见的疾病,引起下腰痛,放射状的腿部疼痛,和感觉障碍。临床前研究在很大程度上依赖于人类疾病的标准化动物模型来预测临床治疗效果以及识别和研究人类受试者中的潜在不良事件。目前制备LDH模型的方法包括从自体尾骨盘采集髓核(NP),并应用于相应背根神经节近端的腰神经根。然而,这种手术方法产生了一个模型,该模型显示出与人类观察到的椎间盘突出症非常不同的特征。
为了产生更类似于人类椎间盘突出症的大鼠LDH模型,以及使用白介素-1β(IL-1β)的标准化和统一的LDH模型。
实验性大鼠LDH模型。
我们在没有神经压迫的情况下,通过半椎板切除术将L5-6椎间盘背外侧暴露在右侧。通过用30号针在4mm的深度刺穿暴露的盘开始疝。同时注射白细胞介素-1β(IL-1β)以增强椎间盘退变的病理过程,包括炎症反应,矩阵破坏,和NP的突出。我们进行组织学染色以评估形态学变化,免疫组化分析L5-6椎间盘穿刺部位内和周围炎症和疼痛相关的表达,和实时聚合酶链反应来检查退化过程标记物的表达。此外,我们对大鼠进行了运动测试。
我们发现,与对照组(仅穿刺)相比,IL-1β组显示纤维环和髓核之间的边界严重中断。And,在LDH模型中,IL-1β的注射以更一致的方式导致椎间盘退变和炎症加速。在暴露的椎间盘中穿刺和注射IL-1β始终引起功能缺陷。
本文提出的方法可用作通过注射的IL-1β浓度与手术诱导的疝同时控制椎间盘退变和炎症严重程度的指标。
我们提出的模型可以促进药物开发研究,以评估椎间盘突出症和神经性疼痛的潜在治疗药物的疗效,也可以用于非临床研究,以根据椎间盘退变的严重程度更准确地评估各种治疗策略的有效性。
Lumbar intervertebral disc herniation (LDH) is a common disease that causes low back pain, radiating leg pain, and sensory impairment. Preclinical studies rely heavily upon standardized animal models of human diseases to predict clinical treatment efficacy and to identify and investigate potential adverse events in human subjects. The current method for making the LDH model involves harvesting the nucleus pulposus (NP) from autologous coccygeal discs and applying to the lumbar nerve roots just proximal to the corresponding dorsal root ganglion. However, this surgical method generates a model that exhibits very different characteristics of disc herniation than that observed in human.
To produce a rat LDH model that better resembles disc herniation in humans and a standardized and uniform LDH model using Interleukin-1 beta (IL-1β).
Experimental rat LDH model.
We exposed the L5-6 disc dorsolaterally on the right side through hemi-laminectomy without nerve compression. Herniation was initiated by puncturing the exposed disc with a 30-gauge needle at a depth of 4 mm. Interleukin-1 beta (IL-1β) was injected simultaneously to heighten the pathological processes of disc degeneration, including inflammatory responses, matrix destruction, and herniation of the NP. We performed histological staining to assess morphological changes, immunohistochemistry to analyze inflammation- and pain-related expression within and around the puncture site of the L5-6 disc, and real-time polymerase chain reaction to examine expression of markers for degenerative processes. In addition, we performed locomotor tests on the rats.
We found that the IL-1β groups showed that the border between the annulus fibrosis and nucleus pulposus was severely interrupted compared to that of the control (puncture only) group. And, the injection of IL-1β leads to accelerated disc degeneration and inflammation in a more consistent manner in LDH model. Functional deficit was consistently induced by puncturing and injection of IL-1β in the exposed disc.
The method proposed here can be used as an index to control the severity of disc degeneration and inflammation through the injected IL-1β concentration concurrent with surgically induced herniation.
Our proposed model may facilitate research in drug development to evaluate the efficacy of potential therapeutic agents for disc herniation and neuropathic pain and may also be used for nonclinical studies to more accurately assess the effectiveness of various treatment strategies according to the severity of disc degeneration.