| 张洪跃,周潘宇,汪 洋,许硕贵,王秀会.白细胞介素-17对体外培养髓核细胞增殖和代谢的影响[J].现代生物医学进展英文版,2017,17(12):2218-2222. |
| 白细胞介素-17对体外培养髓核细胞增殖和代谢的影响 |
| The Effect of IL-17 on the Proliferation and Metabolism of Nucleus Pulposus Cells Cultured in Vitro |
| Received:January 10, 2017 Revised:February 08, 2017 |
| DOI：10.13241/j.cnki.pmb.2017.12.005 |
| 中文关键词: 白细胞介素-17 细胞因子 炎症 椎间盘退变 |
| 英文关键词: Interleukin-17 Cytokine Inflammation Intervertebral disc degeneration |
| 基金项目:上海市卫生局科研计划项目（20134394） |
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| 中文摘要: |
| 摘要 目的：探究白细胞介素-17（interleukin-17，IL-17）对体外培养髓核细胞增殖和细胞代谢的影响。方法：髓核细胞取自经核磁共振影像确认需手术的退变椎间盘组织，建立体外培养体系。用2、5、10、15、20 ng/mL IL-17刺激髓核细胞72 h后，MTS法检测细胞增殖情况。用适当浓度IL-17刺激细胞48 h或96 h后，采用实时定量-PCR和免疫印迹方法检测基质和组织代谢相关基因的mRNA和蛋白表达。结果：IL-17刺激可以抑制体外培养髓核细胞的增殖，且15 ng/mL浓度的抑制作用最强。15 ng/mL IL-17刺激髓核细胞后，聚集蛋白聚糖（aggrecan ，ACAN）和I型胶原（type I collagen，COL1A1）mRNA表达水平显著下降(P<0.05)，基质金属蛋白酶（matrix metalloproteinase-3，MMP3）、金属蛋白酶3组织抑制剂（tissue inhibitor of metalloproteinase-3，TIMP3）的mRNA表达水平显著上升(P<0.05)。COL2A1 mRNA的表达下降，MMP13、含Ⅰ型血小板结合蛋白基序的结聚蛋白样金属蛋白酶（a disintegrin like and metalloproteinase with thrombospondin type Ⅰmotifs-4，ADAMTS4)、ADAMTS5、TIMP1 mRNA的表达上升，但差异均不显著（P>0.05）。IL-17刺激48 h时，COL1A1的蛋白水平明显下降（P=0.010），而ADAMTS5的蛋白水平显著上升（P=0.005）。但刺激96h时，COL1A1的蛋白表达下降，ADAMTS5的蛋白表达上升，但无显著差异（P>0.05）；COL2A1的蛋白表达水平显著下降（P=0.037）。结论：IL-17可抑制体外培养髓核细胞的增殖及代谢，在椎间盘的退变过程中可能发挥了重要的促进作用。 |
| 英文摘要: |
| ABSTRACT Objective: To investigate the role of interleukin-17 (interleukin-17, IL-17) on proliferation and metabolism of nucleus pulposus cells cultured in vitro. Methods: The magnetic resonance imaging showed that nucleus pulposus cells were isolated from human degenerative disc tissues and cultured in vitro. Cells were cultured without or with different concentrations of IL-17. After 72 hours of stimulation by IL-17 of 2, 5, 10, 15, 20 ng/mL, the rate of proliferation inhibition was measured by MTS; Nucleus pulposus cells were cultured without or with an proper concentration of IL-17 for 48 or 96 hours. We used Real-time PCR and Western Blot method to measure the mRNA expression and protein expression levels of matrix macromolecules and tissue degradation genes. Results: It reported that the proliferation of nucleus pulposus cells cultured in vitro was inhibited with the stimulation of IL-17, while the inhibition effect of 15ng/mL IL-17 was significantly stronger. At the dose of 15 ng/mL, the stimulation of IL-17 contributed to multiple cellular responses, including increased mRNA expression of aggrecan (aggrecan, ACAN) and type I collagen (type I collagen, COL1A1) genes (P<0.05), and significantly decreased mRNA expression of tissue degradation genes, matrix metalloproteinase-3 (matrix metalloproteinase-3，MMP3)and tissue inhibitor of metalloproteinase-3 (tissue inhibitor of metalloproteinase-3, TIMP3) (P<0.05). The mRNA expression of COL2A1 and MMP13, a disintegrin like and metalloproteinase with thrombospondin type Ⅰmotifs-4 (a disintegrin like and metalloproteinase with thrombospondin type Ⅰmotifs-4, ADAMTS4), ADAMTS5, TIMP1 genes was increased, but the difference was not significant (P> 0.05). After 48 hours of stimulation by IL-17, the results of Western Blot showed that the level of COL1A1 was dramaticlly decreased (t=-2.814, P=0.010), however the peptidases (ADAMTS5) level was significantly increased (t=3.131, P=0.005).However, after 48 hours of stimulation by IL-17, the protein expression of COL1A1 decreased and the protein expression of ADAMTS5 increased, but there was no significant difference (P>0.05); COL2A1 protein expression level was significantly decreased (P=0.037). Conclusion: These findings suggest that IL-17 can inhibit the proliferation of nucleus pulposus cells in vitro and affect its metabolism, which leads to the degenerative changes of disc disease. |
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