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| LUO Junnan,GUO Dongming,WEN Shifeng.Expression of melatonin receptors in lumbar nucleus pulposus with degeneration of different grades[J].Chinese Journal of Spine and Spinal Cord,2018,(9):839-845. |
| Expression of melatonin receptors in lumbar nucleus pulposus with degeneration of different grades |
| Received:January 12, 2018 Revised:June 28, 2018 |
| English Keywords:Melatonin receptors Nuclear transcription factor -κB Degeneration Lumbar Nucleus pulposus |
| Fund:广州市海珠区科技计划项目（201304231038742045）；广东省科技计划项目（编号：2011A032100001） |
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| English Abstract: |
| 【Abstract】 Objectives: To investigate the expression of melatonin receptor type 1(MT1), melatonin receptor type 2(MT2) and nuclear transcription factor-κB(NF-κB) in lumbar nucleus pulposus with degeneration of different grades. Methods: Thirty-one patients enrolled in our hospital from March 2014 to Janurary 2016 were selected(19 males and 12 females, age 47-62, mean age 56.9±4.9). Magnetic resonance imaging was used to identify the grade of intervertebral disc degeneration with Pfirrmann grading system. Then the nucleus pulposus samples were collected during surgery. 11 samples of grade Ⅳ were allocated into degeneration Ⅰ group; 12 samples of grade V into degeneration Ⅱ group; 8 samples lower than grade Ⅲ into control group. MRNA expressions of MT1, MT2, p65 NF-κB and phosphorylated p65(p-p65) NF-κB were examined by real-time quantitative PCR. Protein expressions of MT1, MT2, p65 NF-κB were also evaluated by immunoblot and immunohistochemistry. Correlation between MT1, MT2 and NF-κB was analyzed by using Spearman method. Results: The results of qRT-PCR showed that MT1 and MT2 mRNA expression in control group were 1.00±0.04 and 1.00±0.04, respectively, while the expressions of MT1 mRNA in degeneration I and degeneration Ⅱ group were 0.68±0.06 and 0.52±0.03. MT2 mRNA expressions were 0.77±0.04 and 0.68±0.06. The difference among these three groups was significant(P<0.05). NF-κB mRNA expressions in three groups were 1.00±0.03, 0.99±0.07 and 0.98±0.05, with insignificant difference(P＞0.05). Western blot results showed that MT1 and MT2 protein levels in control group were 1.00±0.05 and 1.00±0.05, while MT1 protein levels in degeneration I and degeneration Ⅱ group were 0.73±0.07 and 0.56±0.04. MT2 protein levels in degeneration I and degeneration Ⅱ group were 0.62±0.05 and 0.55±0.04. The difference among these three groups was significant(P<0.05). P65 protein levels in three groups were 1.00±0.05, 1.00±0.06 and 0.99±0.04, with insignificant difference(P=0.920). Phosphorylated p65 (p-p65) protein level in control group was 1.00±0.07, while those in degeneration I and degeneration Ⅱ group were 1.43±0.08 and 1.49±0.08, the difference among these three groups was also significant(P<0.05). The results of immunohistochemistry showed that MT1 and MT2 in degenerated nucleus pulposus were inversely correlated with the expression of p-p65 protein (P<0.05). The correlation coefficients were -0.6492 and -0.6190. Conclusions: With the increase of degeneration grade in human nucleus pulposus, the expression of melatonin receptors is suppressed and the activity of NF-κB is promoted. |
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