基质细胞衍生因子-1在人正常和退变椎间盘中的表达、分布及意义

he expression, distribution and significance of stromal cell-derived factor-1 in the normal and degenerative intervertebral disc

中文关键词:  基质细胞衍生因子-1  椎间盘退变  正常椎间盘

英文关键词:Stromal cell-derived factor-1  Intervertebral disc degeneration  Normal intervertebral disc

中文摘要:

  【摘要】 目的：比较人正常和退变椎间盘髓核中央区、髓核与纤维环交界区、纤维环外层、纤维环内层及软骨终板5个相邻部位的基质细胞衍生因子-1（SDF-1）的表达情况，探讨退变椎间盘SDF-1的表达在上述五个相邻部位是否具有差异。方法：获取20例腰椎间盘突出症患者的椎间盘（L4/5，病理证实为退变椎间盘组织），并纳入退变组；7例急性腰椎爆裂骨折患者的椎间盘（L3/4）及3例脊柱侧凸患者的椎间盘（L2/3，病理证实为正常椎间盘组织），纳入正常组。HE染色观察两组椎间盘的组织学结构，免疫组织化学、Western-blot、rt-PCR检测椎间盘髓核中央区、髓核与纤维环交界区、纤维环内层、纤维环外层及软骨终板5个部位的SDF-1表达情况及分布，并将退变椎间盘上述5个部位的SDF-1免疫组化积分光密度值与供者年龄、椎间盘Thompson分级作相关分析。结果：正常组的上述5个部位SDF-1免疫组化积分光密度值分别为420.87±89.93、407.80±100.76、380.02±85.05、342.89±63.76、344.06±88.97；Western-blot灰度比值分别为0.08±0.03、0.08±0.05、0.07±0.04、0.05±0.03、0.05±0.02；rt-PCR灰度比值分别为0.22±0.06、0.22±0.05、0.20±0.04、0.20±0.04、0.20±0.04。退变组上述5个部位SDF-1免疫组化积分光密度值分别为7355.13±2271.74、5959.58±2436.23、5397.49±3044.80、1605.44±825.31、361.91±104.22；Western-blot灰度比值分别为0.55±0.29、0.52±0.28、0.42±0.18、0.21±0.12、0.06±0.04；rt-PCR灰度比值分别为0.75±0.30、0.71±0.26、0.55±0.22、0.36±0.17、0.22±0.13。退变椎间盘5个部位的SDF-1表达均高于正常组（P<0.05），且SDF-1表达量髓核中央区＞纤维环内层＞纤维环外层＞软骨终板，差异有统计学意义（P<0.05），而髓核中央区和髓核与纤维环交界区比较无统计学差异（P>0.05）；退变椎间盘上述5个部位的SDF-1表达与椎间盘Thompson分级、年龄呈正相关性（P<0.05）。结论：SDF-1广泛表达于退变和正常的椎间盘中，其表达在退变椎间盘中上述5个相邻部位存在差异。

英文摘要:

  【Abstract】 Objectives: To compare the expressions of stromal cell-derived factor-1(SDF-1) in different areas of normal and degenerative intervertebral disc, and to explore the expression pattern of SDF-1 in five adjacent parts of the intervertebral disc, which was the inner nucleus pulposus(INP), transitional nucleus pulposus(TNP), inner annulus fibrous(IAF), outer annulus fibrous(OAF) and cartilage endplate(CEP). Methods: 20 patients with lumbar intervertebral disc herniation were collected as the degeneration group(the degenerated discs were all located in L4/5, confirmed by surgical pathology). 7 patients with acute bursting fracture and 3 patients with scoliosis were included as the control group. Intervertebral disc was divided into five areas, which included INP, TNP, IAF, OAF, CEP. HE staining was used to detect the structure of the intervertebral disc in both groups; immunohistochemistry, Western blot and rt-PCR were used to detect the expression and distribution of SDF-1 in the five adjacent areas between two groups. Finally, the relationship between the results of immunohistochemistry of SDF-1, donor age and Thompson classification of the intervertebral disc in both groups were analyzed. Results: In control group, the immunohistochemistry IOD value of SDF-1 in the above-mentioned five areas was 420.87±89.93, 407.80±100.76, 380.02±85.05, 342.89±63.76, 344.06±88.97 respectively; Gray value of Western-blot was 0.08±0.03, 0.08±0.05, 0.07±0.04, 0.05±0.03, 0.05±0.02 respectively; Gray value of rt-PCR was 0.22±0.06, 0.22±0.05, 0.20±0.04, 0.20±0.04, 0.20±0.04 respectively. As for the degeneration group, the OD value of SDF-1 in those five areas was 7355.13±2271.74, 5959.58±2436.23, 5397.49±3044.80, 1605.44±825.31, 361.91±104.22 respectively; Gray value of Western-blot was 0.55±0.29, 0.52±0.28, 0.42±0.18, 0.21±0.12, 0.06±0.04 respectively; Gray value of rt-PCR was 0.75±0.30, 0.71±0.26, 0.55±0.22, 0.36±0.17, 0.22±0.13 respectively. The expression of SDF-1 in the five areas in the degeneration group was higher than the control group(P<0.05). Besides, the expression of SDF-1 in the five areas presented as a gradient manner, with the highest in the inner annulus fibrosus and the lowest in the cartilage endplate, which showed statistically significant difference(P<0.05). However, the central nucleus pulposus and border area nucleus pulposus had no difference(P>0.05). The expressions of SDF-1 in five areas had a positive correlation with the donor age and the Thompson classification. Conclusions: SDF-1 is expressed widely in the degenerative and normal intervertebral disc, and the distribution of SDF-1 in degenerated intervertebral disc at adjacent areas was in a gradient manner.