张雯，徐辉，罗大伟，冯洪永，纪昌宾，杨克石，李彬，李昆朋.牵张应力对退变椎间盘髓核细胞增殖凋亡及细胞外基质表达的影响[J].中华物理医学与康复杂志,2018,40(10):721-726
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| 牵张应力对退变椎间盘髓核细胞增殖凋亡及细胞外基质表达的影响 |
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| DOI： |
| 中文关键词: 牵张应力 退变髓核细胞 增殖 凋亡 细胞外基质 |
| 英文关键词: Mechanical stress Stretching Degenerate nuclei Pulposus cells Proliferation Apoptosis Extracellular matrices |
| 基金项目:山东省自然科学基金资助项目(ZR2014HL027、ZR2017MH112)；山东省医药卫生科技发展计划项目（2016WS0206） |
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| 中文摘要: |
| 目的 探讨不同强度牵张应力刺激对体外培养的人退变髓核细胞增殖凋亡和细胞外基质表达的影响。 方法 分选并体外培养人退变的髓核细胞，使用四点弯曲细胞力学装置对细胞施加不同强度的牵张应力，根据施加牵张应力强度的不同分为对照组（未给予应力刺激）、低强度组（1000 μ）、中强度组（2000 μ）和高强度组（4000 μ）四组。采用流式细胞仪测定退变髓核细胞的增殖情况；采用实时荧光定量聚合酶链反应（qPCR）检测各组细胞的细胞增殖核抗原(PCNA)、细胞凋亡相关蛋白B淋巴细胞瘤-2基因（Bcl-2）与Bcl-2相关X蛋白（Bax）、以及Ⅱ型胶原（ColⅡ）和蛋白聚糖(Aggrecan)的基因表达情况并进行分析。 结果 在不同强度的牵张应力作用下，退变髓核细胞的增殖凋亡和细胞外基质分泌呈现不同的变化。随着施加力学强度的增加，髓核细胞的增殖指数和PCNA基因表达水平先升高而后又逐渐降低，差异有统计学意义（P<0.05）。髓核细胞凋亡相关基因Bcl-2/Bax mRNA值在1000 μ强度牵张应力作用下为对照组的1.53倍，而在2000 μ和4000 μ强度下分别为0.71和0.43，同样呈现出随应力刺激增加先升高又降低的趋势，差异均有统计学意义（P<0.05）。给予1000 μ强度应力刺激后，Col Ⅱ和Aggrecan均有不同程度的表达增加，分别增加了2.1倍和2.3倍，与对照组比较,差异有统计学意义（P<0.05）。随着牵张应力强度的增加，Col Ⅱ和Aggrecan基因表达逐渐降低，在4000 μ强度牵张应力刺激时，二者基因表达最低，差异具有统计学意义（P<0.05）。 结论 不同强度的牵张应力对人退变髓核细胞的增殖凋亡以及细胞外基质的表达作用不同。 |
| 英文摘要: |
| Objective To observe in vitro the effect of mechanical stress at different intensities on the proliferation, apoptosis and extracellular matrix of degenerative human nucleus pulposus cells. Methods The cells were isolated and cultured in vitro, and divided into a control group, a low-intensity group, a medium-intensity group and a high-intensity group. The low-, medium- and high-intensity groups were stretched mechanically by 1000 μ, 2000 μ or 4000 μ respectively for 6 hours using a four-point bending system, while the control group was not stressed. Flow cytometry was used to explore any changes in the cell cycle and the proliferation index (PI). The expression of proliferating cell nuclear antigen (PCNA), B-cell lymphoma-2 (BCL-2)/Bax, collagen II and aggrecan were measured using real-time quantitative polymerase chain reactions. Results The mechanical stretching significantly influenced proliferation, apoptosis and the extracellular matrices compared with the control group. The PI and PCNA expression increased at first and then decreased gradually with the exercise intensity. Compared with the control group, the mRNA expression level of Bcl-2/Bax increased significantly to 1.53 times that of the control group after 1000 μ stretching, but to only 0.71 times that of the control group at 2000 μ and 0.43 times at 4000 μ. The gene expression of collagen II increased significantly by 1.1 times and that of aggrecan by 1.3 times after 1000 μ stress stimulation compared with the control group (P≤0.05). However, the expression of collagen II and aggrecan was inhibited significantly at 2000 and 4000 μ , with the lowest levels at 4000 μ (P≤0.05). Conclusion Stretching at different intensities has different effects on the proliferation, apoptosis and extracellular matrix of human pulposus cells with degenerate nuclei. |
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