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| YANG Kuang,LI Haiyin,ZHANG Wei.Expression of sphingosine 1-phosphate receptors in degenerative nucleus pulposus with different degrees[J].Chinese Journal of Spine and Spinal Cord,2015,(12):1083-1089. |
| Expression of sphingosine 1-phosphate receptors in degenerative nucleus pulposus with different degrees |
| Received:August 25, 2015 Revised:October 14, 2015 |
| English Keywords:Intervertebal disk degeneration Sphingosine 1-phosphate receptors Nucleus pulposus cells |
| Fund:国家自然科学基金资助项目（81572208） |
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| English Abstract: |
| 【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. |
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