杨 匡,李海音,张 伟,路 康,周 跃,李长青.1-磷酸鞘氨醇受体在不同退变程度髓核组织中的表达变化[J].中国脊柱脊髓杂志,2015,(12):1083-1089. |
1-磷酸鞘氨醇受体在不同退变程度髓核组织中的表达变化 |
中文关键词: 椎间盘退变 1-磷酸鞘氨醇受体 髓核细胞 |
中文摘要: |
【摘要】 目的:比较不同退变程度人椎间盘髓核组织中3种1-磷酸鞘氨醇受体(S1PR1/2/3)表达水平的差异,探讨椎间盘中S1PR表达水平与椎间盘退变的关系。方法:收集腰椎间盘退行性病变患者手术切除的椎间盘组织,其中轻度退变(Pfirrmann分级Ⅳ级)22例,严重退变(Pfirrmann分级Ⅴ级)14例;同时取6例无椎间盘退变患者(单纯腰椎椎体骨折,Pfirrmann分级Ⅱ级)手术切除的椎间盘组织作为对照组;通过HE染色以及Saf-O染色观察不同退变程度椎间盘的组织学变化,免疫组化检测不同退变程度组织中的S1PR表达水平;Ⅱ型胶原酶消化分离提取原代髓核细胞,通过Real-time PCR、Western-bolt检测不同退变程度椎间盘髓核细胞中S1PR的表达水平,并通过细胞免疫化学方法对S1PR进行定位。结果:HE染色及Saf-O染色结果显示退变椎间盘的纤维环出现破损,髓核细胞形成明显的集落,细胞外基质减少。免疫组化结果显示正常和轻度退变的髓核组织中3种受体(S1PR1/2/3)都有表达,严重退变的组织中表达极弱;Real-time PCR结果显示对照组髓核细胞中 S1PR1/2/3 的mRNA表达水平分别是严重退变组的5.34±0.52倍、7.25±0.04倍、1.92±0.06倍,轻度退变组S1PR1/2/3 的mRNA表达水平分别是严重退变组的4.35±2.45倍、4.96±3.44倍、2.19±0.82倍;Western-blot发现对照组和轻度退变组髓核细胞中S1PR1/2/3均有表达,严重退变组表达水平较低;免疫细胞化学显示S1PR主要集中在髓核细胞的细胞质和细胞膜上。结论:髓核组织中主要表达S1PR1/2/3,在严重退变的髓核组织和细胞中其表达水平明显下降,S1P及其受体可能参与椎间盘髓核组织的退变过程。 |
Expression of sphingosine 1-phosphate receptors in degenerative nucleus pulposus with different degrees |
英文关键词:Intervertebal disk degeneration Sphingosine 1-phosphate receptors Nucleus pulposus cells |
英文摘要: |
【Abstract】 Objectives: To compare the expression of sphingosine 1-phosphate receptors(S1PRs) in degenerative nucleus pulposus with different degrees, and to analyze the relationship between the expression level and the degeneration. Methods: 22 mild degenerative intervertebral discs(Pfirrmann grade Ⅳ) and 14 advanced degenerative(Pfirrmann grade V) discs were collected as samples, 6 normal samples(Pfirrmann grade Ⅱ, simple vertebral fracture) were used as controls. HE staining and Saf-O staining was used to show the histological changes of degeneration. The expression of S1PRs was detected by immunohistochemical method. Primary nucleus pulposus cells was obtained by collagenas Ⅱ digestion. Real-time PCR and Western-blot were used to assess the S1PRs in the nucleus pulposus cells. The cytochemistry was used to evaluate the location of S1PRs in nucleus pulposus cells. Results: HE and Saf-O staining showed intervertebral disk with advanced degeneration: fracture appeared in annulus fibrosus, cells bind together into clones in nucleus pulposus and extracellular matrix. The immunohistochemical result showed that normal and mild degenerated nucleus pulposus expressed S1PR1/2/3, almost not in the advanced degenerated cells. The real-time PCR showed that the expression of S1PR decreases in advanced degenerated nucleus pulposus(the normal to the advanced degenerated was 5.34±0.52-flod, 7.25±0.04-flod, 1.92±0.06-flod, the mild degenerated to the advanced was 4.35±2.45-flod, 4.96±3.44-flod, 2.19±0.82-flod). Western-blot result showed that NP cells form control and mild degeneration expressed S1PR1/2/3, while much weak in the advanced degenerated NP cells. The cytochemistry showed that S1PRs focused in the membranes and cytoplasm of cells. Conclusions: The intervertebral disc mainly expresses S1PR1/2/3, and the expression decreases in the advanced degenerative nucleus pulposus. The S1P and its receptor may play a role in the degeneration of intervertebral disc. |
投稿时间:2015-08-25 修订日期:2015-10-14 |
DOI: |
基金项目:国家自然科学基金资助项目(81572208) |
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