中国组织工程研究

中国组织工程研究 ›› 2022, Vol. 26 ›› Issue (12): 1816-1821.doi: 10.12307/2022.501

• 骨与关节生物力学 bone and joint biomechanics • 上一篇    下一篇

力学因素对椎间盘退行性变的影响及机制

尹逊路1，金哲峰1，朱立国1，2，3，冯敏山1，2，3，于  杰1，魏  戌1，展嘉文1，高春雨1，银  河1，梁  龙4，韩  涛1，孙  凯1，谢  瑞1   

  1. 1中国中医科学院望京医院，北京市   100102；2北京市中西医结合骨伤研究所，北京市   100700；3中医正骨技术北京市重点实验室，北京市   100700；4安徽中医药大学第一附属医院，安徽省合肥市   230031
  • 收稿日期:2021-06-02 修回日期:2021-06-17 接受日期:2021-07-10 出版日期:2022-04-28 发布日期:2021-12-14
  • 通讯作者: 金哲峰，硕士，副主任医师，中国中医科学院望京医院，北京市 100102 冯敏山，博士，主任医师，中国中医科学院望京医院，北京市 100102；北京市中西医结合骨伤研究所，北京市 100700；中医正骨技术北京市重点实验室，北京市 100700
  • 作者简介:尹逊路，男，1988年生，山东省肥城市人，汉族，2019年中国中医科学院毕业，博士，主治医师，主要从事脊柱与相关疾病研究。 朱立国，男，1961年生，黑龙江省牡丹江市人，汉族，2011年天津中医药大学毕业，博士，主任医师，主要从事脊柱与相关疾病研究。
  • 基金资助:
    国家自然科学基金面上项目(81774330)，项目负责人：冯敏山；北京市自然科学基金青年项目(7214297)，项目负责人：尹逊路

Effect and mechanism of mechanical factors on intervertebral disc degeneration

Yin Xunlu1, Jin Zhefeng1, Zhu Liguo1, 2, 3, Feng Minshan1, 2, 3, Yu Jie1, Wei Xu1, Zhan Jiawen1, Gao Chunyu1, Yin He1, Liang Long4, Han Tao1, Sun Kai1, Xie Rui1    

  1. 1Wangjing Hospital, Chinese Academy of Traditional Chinese Medicine, Beijing 100102, China; 2Beijing Institute of Integrated Traditional Chinese and Western Medicine, Beijing 100700, China; 3Beijing Key Laboratory of Bone Setting Technology of Traditional Chinese Medicine, Beijing 100700, China; 4First Affiliated Hospital of Anhui University of Traditional Chinese Medicine, Hefei 230031, Anhui Province, China
  • Received:2021-06-02 Revised:2021-06-17 Accepted:2021-07-10 Online:2022-04-28 Published:2021-12-14
  • Contact: Jin Zhefeng, Master, Associte chief physician, Wangjing Hospital, Chinese Academy of Traditional Chinese Medicine, Beijing 100102, China Feng Minshan, MD, Chief physician, Wangjing Hospital, Chinese Academy of Traditional Chinese Medicine, Beijing 100102, China; Beijing Institute of Integrated Traditional Chinese and Western Medicine, Beijing 100700, China; Beijing Key Laboratory of Bone Setting Technology of Traditional Chinese Medicine, Beijing 100700, China
  • About author:Yin Xunlu, MD, Attending physician, Wangjing Hospital, Chinese Academy of Traditional Chinese Medicine, Beijing 100102, China Zhu Liguo, MD, Chief physician, Wangjing Hospital, Chinese Academy of Traditional Chinese Medicine, Beijing 100102, China; Beijing Institute of Integrated Traditional Chinese and Western Medicine, Beijing 100700, China; Beijing Key Laboratory of Bone Setting Technology of Traditional Chinese Medicine, Beijing 100700, China Yin Xunlu and Zhu Liguo contributed equally to this article.
  • Supported by:
    the National Natural Science Foundation of China, No. 81774330 (to FMS); Beijing Natural Science Foundation (Youth Project), No. 7214297 (to YXL)

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摘要：

文题释义：
脊柱运动节段：维持脊柱稳定性的基本功能单位是运动节段，包括相邻的两节脊柱及其间的椎间盘、关节突关节以及韧带结构等。
椎间盘退变：指椎间盘的生物化学成分和结构发生改变，导致椎间盘生物力学功能下降的病理学过程。

背景：课题组前期研究发现，力学刺激可影响Wnt/β-catenin信号通路中多个关键蛋白(包括β-catenin、GSK-3β蛋白等)，但其是否通过调控髓核细胞中Wnt/β-catenin信号通路有待于进一步明确。
目的：阐明持续负载压力诱导椎间盘退行性变的效应特点，探讨力学因素介导Wnt/β-catenin信号通路对椎间盘退行性变的影响机制。
方法：获取70只新西兰大白兔的脊柱运动节段，采用自主研发的离体兔脊柱运动节段模型，根据不同大小的持续负载压力将其分为空白对照组(0 kg)、低压力组(0.5 kg)、中压力组(1 kg)、高压力组(3 kg)；再选取最优持续负载压力组(对照组)，分别采用Wnt/β-catenin信号通路特异性激活剂SB216763(激活剂组)及抑制剂ICG001(抑制剂组)干预，连续干预3 d。采用苏木精-伊红染色、DAB染色、Western blot、RT-PCR等技术观察髓核细胞和功能。实验方案经中国中医科学院望京医院医学伦理委员会批准。
结果与结论：①中压力(1 kg)下髓核细胞存活率较高；3 d时持续负载压力可明显诱导椎间盘退行性变发生；②与对照组相比，激活剂组中髓核组织内髓核细胞数量明显增多、细胞形态规则、髓核中纤维组织和瘢痕不明显；β-catenin及聚集蛋白聚糖蛋白表达水平较高、GSK-3β蛋白表达水平显著降低(P < 0.05)，聚集蛋白聚糖及Ⅱ型胶原蛋白 mRNA表达水平显著升高(P < 0.05)；③与对照组相比，抑制剂组髓核组织内髓核细胞明显减少、细胞形态多成梭形、髓核中纤维组织及瘢痕较明显；GSK-3β及β-catenin表达量差异无显著性意义，聚集蛋白聚糖及Ⅱ型胶原蛋白 mRNA表达水平显著降低(P < 0.05)；④结果说明，持续负载力可通过介导Wnt/β-catenin信号通路调控髓核细胞影响椎间盘退行性变的进程。

https://orcid.org/0000-0002-3388-4215 (尹逊路)；https://orcid.org/0000-0002-1931-8647(朱立国) 

中国组织工程研究杂志出版内容重点：人工关节；骨植入物；脊柱；骨折；内固定；数字化骨科；组织工程


关键词: 腰椎间盘突出症, 椎间盘退行性变, 持续负载压力, 髓核细胞, Wnt/β-catenin信号通路

Abstract: BACKGROUND: Our previous study found that mechanical stimulation can affect many key proteins in the Wnt/β-catenin signaling pathway (including β-catenin and GSK-3β protein), but whether it regulates the Wnt/β-catenin signaling pathway in nucleus pulposus cells remains to be clarified.  
OBJECTIVE: To clarify the characteristics and effect of continuous loading pressure on intervertebral disc degeneration, and explore the mechanism of mechanical factors-mediated Wnt/β-catenin signaling pathway on intervertebral disc degeneration.
METHODS:  Seventy New Zealand white rabbits were selected to establish a rabbit spinal motion segment model, which were randomly divided into blank control group (0 kg), low pressure group (0.5 kg), medium pressure group (1 kg), and high pressure group (3 kg) according to the sustained loading pressure. Others were divided into control group by using optimal sustained loading pressure, activator group and inhibitor group by using Wnt/β-catenin specific activators (SB216763) and inhibitors (ICG001), for 3 consecutive days. Hematoxylin-eosin staining, DAB staining, western blot assay, and RT-PCR were used to observe the function of nucleus pulposus cells. The protocols were approved by Medical Ethics Committee of Wangjing Hospital, China Academy of Chinese Medical Sciences.  
RESULTS AND CONCLUSION: (1) The survival rate of nucleus pulposus cells was high under medium pressure (1 kg). Sustained loading pressure could induce intervertebral disc degeneration at 3 days. (2) Compared with the control group, the number of nucleus pulposus cells in the activator group was significantly increased; the cell morphology was regular, and the fibrous tissue and scar in the nucleus pulposus were not obvious. The expression levels of β-catenin and Aggrecan protein were higher and the experssion of GSK-3β protein was significantly decreased (P < 0.05). The expression levels of Aggrecan and Collagen II mRNA were significantly increased (P < 0.05). (3) Compared with the control group, the number of nucleus pulposus cells in the inhibitor group was significantly reduced; the cell morphology was fusiform, and the fibrous tissue and scar in the nucleus pulposus were more obvious; the expression levels of GSK-3β and β-catenin were not significantly different, but the expression of Aggrecan and Collagen II mRNA decreased significantly (P < 0.05). (4) It is concluded that sustained loading force regulates the process of intervertebral disc degeneration in nucleus pulposus cells by mediating Wnt/β-catenin signaling pathway.

Key words: lumbar disc herniation, intervertebral disc degeneration, continuous load pressure, nucleus pulposus cells, Wnt/β-catenin signaling pathway

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