三种针刺注射无水乙醇法构建椎间盘退变大鼠模型的比较

Comparative study of three needle injection of ethanol methods for establishing intervertebral disc degeneration models in rat

中文关键词:  椎间盘退变  针刺注射无水乙醇  大鼠  椎间高度指数

英文关键词:Intervertebral disc degeneration  Percutaneous injection of anhydrous ethanol  Rat  Disc height index

中文摘要:

  【摘要】 目的：比较针刺髓核、纤维环、终板注射无水乙醇所构建三种大鼠椎间盘退变（intervertebral disc degeneration，IDD）模型的差异。方法：将45只6周龄SD大鼠采用随机数字表法随机分为三组，每组15只，分别采用髓核注射（髓核注射组）、纤维环注射（纤维环注射组）以及终板注射（终板注射组）三种针刺注射无水乙醇法对大鼠Co6/7椎间盘构建IDD模型，在造模术后1、2、3、4周采用随机数字表法分别在三组各随机挑选3只大鼠进行X线透视检查，并测量计算大鼠Co6/7的椎间高度指数（disc height index，DHI），随后进行过量麻醉处死取下Co6/7椎间盘连同相邻椎体进行苏木精-伊红（hematoxylin-eosin，HE）染色，观察退变情况并对退变程度进行组织学评分。结果：髓核注射组、纤维环注射组各有1只大鼠因麻醉意外死亡；终板注射组有2只大鼠因术后感染死亡。造模术后1、2、3、4周在各组剩余大鼠中每组选取3只，共36只大鼠纳入结果分析，剩余大鼠通过过量麻醉处死。各组造模后1周Co6/7 DHI均较造模前降低（髓核注射组0.117±0.002 vs 0.134±0.005；纤维环注射组0.126±0.005 vs 0.133±0.005；终板注射组0.127±0.002 vs 0.130±0.007，均P<0.05），且随着时间延长均逐渐降低。各组造模后，纤维环断裂，纤维排列逐渐紊乱，髓核基质、细胞逐渐丢失甚至全部消失，椎间盘逐渐坍塌，椎间隙逐渐狭窄。髓核注射组较另外两组退变速度更快，程度更高，造模4周后，髓核注射组DHI为0.090±0.008，低于纤维环注射组（0.106±0.007，P<0.05）及终板注射组（0.109±0.000，P<0.05）；组织学评分髓核注射组为10.00±0.82分，高于纤维环注射组（9.00±1.41分，P<0.05）及终板注射组（8.75±0.96分，P<0.05）。结论：三种针刺注射无水乙醇法构建的三种IDD模型各有其特点，髓核注射法退变最快、程度最高，终板注射法退变最慢，但更符合自然退变过程，纤维环注射法介于两者之间。

英文摘要:

  【Abstract】 Objectives: To compare the differences in three rat intervertebral disc degeneration(IDD) models constructed by needle injections of ethanol into the nucleus pulposus, annulus fibrosus, and endplate, respectively. Methods: 45 6-week-old Sprague-Dawley rats were randomly divided into three groups using a random number table method, with 15 rats in each group. Three different needle injection of ethanol methods including nucleus pulposus injection(nucleus pulposus injection group), annulus fibrosus injection(annulus fibrosus injection group), and endplate injection(endplate injection group) were used to construct IDD models at the Co6/7 intervertebral disc in rats. At 1, 2, 3, and 4 weeks post-modeling, 3 rats from each group were randomly selected using a random number table for X-ray examination to measure and calculate the disc height index(DHI) of the Co6/7 in rats. Subsequently, rats were euthanized under excessive anesthesia, and the Co6/7 intervertebral disc along with adjacent vertebral bodies were harvested for hematoxylin-eosin(HE) staining to observe degeneration, and the degeneration degree was histologically scored. Results: One rat in each of the nucleus pulposus injection group and annulus fibrosus injection group died unexpectedly due to anesthesia； In the endplate injection group, two rats died due to postoperative infection. At 1, 2, 3, and 4 weeks after modeling, 3 rats were selected from each group of the remaining rats, and a total of 36 rats were included in the result analysis. The remaining rats were sacrificed by excessive anesthesia. One week after modeling, the Co6/7 DHI in each group was lower than that before modeling(Nucleus pulposus injection group: 0.117±0.002 vs 0.134±0.005; Annulus fibrosus injection group: 0.126±0.005 vs 0.133±0.005; Endplate injection group: 0.127±0.002 vs 0.130±0.007, all P<0.05), which gradually decreased over time. After modeling in each group, the annulus fibrosus broke, the arrangement of fibers gradually became disordered, the matrix and cells of the nucleus pulposus were gradually lost or even completely disappeared, the intervertebral discs gradually collapsed, and the intervertebral spaces gradually narrowed. The degeneration rate and degree of the nucleus pulposus injection group were faster and higher than those of the other two groups. Four weeks after modeling, the DHI of the nucleus pulposus injection group was 0.090±0.008, which was lower than that of the annulus fibrosus injection group(0.106±0.007, P<0.05) and endplate injection group(0.109±0.000, P<0.05); The histological score of the nucleus pulposus injection group was 10.00±0.82, which was higher than that of the annulus fibrosus injection group(9.00±1.41, P<0.05) and endplate injection group(8.75±0.96, P<0.05). Conclusions: The three needle injection of ethanol methods have their own unique characteristics for constructing IDD models. Nucleus pulposus injection induces the fastest and most severe degeneration, endplate injection leads to slower degeneration but is more in line with natural degeneration processes, and annulus fibrosus injection falls between the other two methods in terms of degeneration characteristics.