雌激素受体在椎间盘组织中的表达及意义

Expression and significance of estrogen receptors in disci intervertebrales tissue

中文关键词:  髓核细胞  雌激素  雌激素受体  免疫组织化学

英文关键词:Nucleus pulposus cell  Estrogen  estrogen receptor  Immunohistochemistry

中文摘要:

  【摘要】 目的：通过观察雌激素α、β受体在正常与退变的人椎间盘组织的表达，探讨雌激素受体（ER）与椎间盘退变的关系。方法：根据改良Pfirrmann分级将收集的椎间盘组织分为三组：对照组，外伤导致腰椎爆裂性骨折手术取出的正常髓核组织（Pfirrmann分级1~2级）；观察组，女性腰椎滑脱及腰椎间盘突出症手术取出的退变髓核组织，Pfirrmann分级3～4级为A组，5级为B组。用HE染色法观察对照组和A、B组各15例人腰椎椎间盘髓核组织及髓核细胞的形态学变化；用免疫组织化学染色法（Elivison二步法）检测对照组和A、B组髓核组织中ER-α、ER-β的表达；采用Western-blot法检测对照组和观察组髓核组织中ER-α、ER-β的表达。结果：HE染色示对照组髓核组织中髓核细胞分布均匀，形态完整，未见明显细胞凋亡现象，细胞外基质染色鲜亮，分布均匀；A、B两组髓核细胞分布不均匀，形态不规则，凋亡现象明显，细胞核增大，染色深，细胞外基质减少，色彩灰暗，B组较A组更为明显。免疫组织化学染色示ER-α、ER-β在对照组的髓核组织见有明显棕黄色颗粒，在A、B两组表达减少，以ER-β减少最显著，经统计学分析，A、B两组与对照组间ER-β的表达有显著性差异（P＜0.01），ER-α的表达较对照组无统计学差异（P＞0.05），其中A组和B组间无明显差异（P＞0.05）。Western-blot方法检测，ER-α表达A组（0.876±0.058）、B组（0.757±0.045）较对照组（0.885±0.036）降低，但无统计学差异（P>0.05）；ER-β表达A组（0.947±0.043）、B组（0.626±0.042）较对照组（1.275±0.150）显著降低（P＜0.01），B组较A组表达明显减少（P＜0.01）。结论：在人的髓核组织中的髓核细胞（类软骨细胞）的胞核和胞浆中均存在ER。ER-α、ER-β在退变的椎间盘组织都明显减少，以ER-β最为明显，提示雌激素可能通过ER-β的介导对髓核细胞功能起调控作用。

英文摘要:

  【Abstract】 Objectives: To investigate the relationship of ER with degeneration of intervertebral disc furtherly by detecting the expression of estrogen receptor in normal and degenerative disc tissues. Methods: According to Pfirrmann grading, dividing into three groups: control group, the nucleus pulposus of the normal lumbar burst fracture; observation group, the nucleus pulposus removed from women who suffered spondylolisthesis and lumbar prominent surgery, according to Pfirrmann grading, divided into group A and B. To observe the morphological changes of nucleus pulposus and nucleus pulposus cells with HE staining in control group and group A, B. To detect the expression and distribution of ER-α, ER-β in nucleus pulposus by immunohistochemical staining(Elivison two-steps) in control group and group A, B. To test the expression of ER-α, ER-β in nucleus pulposus by West-blotting in control group and group A, B. Results: HE staining showed that nucleus pulposus cells in the control group were intact and evenly distributed, no obvious cell apoptosis, extracellular matrix stained bright and evenly distributed. While nucleus pulposus cells in group A, B were irregular shaped and unevenly distributed, Obvious cell apoptosis, the nuclei enlarged, dark staining, extracellular matrix stained gray and unevenly distributed, group B was more obvious than group A. Immunohistochemical staining showed that obvious brown-yellow particles were stained in the nucleus pulposus of the control group, while just a little in the study group, ER-β had been found the most obvious reduced. The statistical analysis between control group and group A, B had significant difference(P<0.01). However, the expression of ER-α did not have any significant difference between the control group and group A, B(P>0.05). There was no significant difference between group A and B(P>0.05). West-blotting showed that compared with control group(0.885±0.036), ER-α in group A(0.876±0.058), group B(0.757±0.045) was lower, but no statistical significance(P>0.05). ER-β in group A(0.947±0.043), group B(0.626±0.042) were significantly reduced than the control group(1.275±0.150)(P<0.01). The expression of ER-β had significant difference between the group A and B(P<0.01). Conclusions: There are expression of estrogen receptors in man′s nucleus pulposus cells. The expression of ER-α, ER-β were significantly reduced in degenerative disc tissue, the ER-β is most obvious. The results suggest that the function of nucleus pulposus cells may be regulated directly by estrogen through ER-β.