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  

The effect of mechanical stretching stress on the proliferation, apoptosis of degenerative disc nucleus pulposus cells and expression of extracellular matrix

作　　者：张雯[1] 徐辉[2] 罗大伟 冯洪永 纪昌宾[2] 杨克石 李彬 李昆朋[2] Zhang Wen;Xu Hui;Luo Dawei;Feng Hongyong;Ji Changbin;Yang Keshi;Li Bin;Li Kunpeng(Department of Ultrasonography,People＇s Hospital of Liaocheng,Liaocheng 252000,China)

机构地区：[1]聊城市人民医院超声医学科,聊城252000 [2]聊城市人民医院骨科,聊城252000

出　　处：《中华物理医学与康复杂志》2018年第10期721-726,共6页Chinese Journal of Physical Medicine and Rehabilitation

基　　金：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）

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（1000μ）、中强度组（2000μ）和高强度组（4000μ）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／BaxmRNA值在1000斗强度牵张应力作用下为对照组的1．53倍，而在2000p和4000斗强度下分别为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汕强度应力刺激后，ColⅡ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，colⅡ和Aggrecan基因表达逐渐降低，在4000斗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。Objective To observe in vitro the effect of mechanical stress at different intensities on the pro- liferation, 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 eell 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 meas- ured 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 2000μ. and 0.43 times at 4000μ. The gene expression of collagen II increased significantly by 1.1 times and that of aggrecan by 1.3 times after 1000μ. stress stimulation compared with the control group （P≤0.05）. However, the expression of collagen Ⅱ and aggrecan was inhibited sig nificantly at 2000 and 4000μ , with the lowest levels at 4000 μ （P ≤ 0.05 ）. Conclusion Stretching at different intensities has different effects on the proliferation, apoptosis and extraeellular matrix of human pulposus cells with degenerate nuclei.

关 键 词：牵张应力 退变髓核细胞 增殖 凋亡 细胞外基质 

分 类 号：R681.53[医药卫生—骨科学]