中国组织工程研究

中国组织工程研究 ›› 2024, Vol. 28 ›› Issue (26): 4170-4177.doi: 10.12307/2024.422

• 肌肉肌腱韧带组织构建 tissue construction of the muscle, tendon and ligament • 上一篇    下一篇

“筋出槽”大鼠模型骨骼肌形态学和功能的特征性变化

杨宗睿1，2，葛海雅1，2，石金玉1，2，汪正明1，2，王媛媛1，2，李正言1，2，杜国庆1，2，詹红生1，2   

  1. 1上海中医药大学附属曙光医院石氏伤科医学中心，上海市  201203；2上海市中医药研究院骨伤科研究所，上海市  201203
  • 收稿日期:2023-05-23 接受日期:2023-07-17 出版日期:2024-09-18 发布日期:2023-09-28
  • 通讯作者: 詹红生，教授，主任医师，博士生导师，上海中医药大学附属曙光医院石氏伤科医学中心，上海市 201203；上海市中医药研究院骨伤科研究所，上海市 201203
  • 作者简介:杨宗睿，男，1996年生，甘肃省平凉市人，汉族，上海中医药大学在读硕士，主要从事慢性筋骨病损的防治研究。
  • 基金资助:
    国家自然科学基金项目(82074466)，项目负责人：詹红生；詹红生全国名老中医药专家传承工作室建设项目(国中医药人教函[2022]75号)，项目负责人：詹红生；上海市慢性筋骨病临床医学研究中心(20MC1920600)，项目负责人：詹红生；上海市临床重点专科“中医骨伤科”(shslczdzk03901)，项目负责人：詹红生；上海高水平地方高校“慢性筋骨病损研究与转化”创新团队(沪教委人[2022]3号)，项目负责人：詹红生

Characteristic changes in morphology and function of skeletal muscles in a rat model of “tendon off-position”

Yang Zongrui1, 2, Ge Haiya1, 2, Shi Jinyu1, 2, Wang Zhengming1, 2, Wang Yuanyuan1, 2, Li Zhengyan1, 2, Du Guoqing1, 2, Zhan Hongsheng1, 2   

  1. 1Shi’s Center of Orthopedics and Traumatology, Shuguang Hospital Affiliated to Shanghai University of Traditional Chinese Medicine, Shanghai 201203, China; 2Institute of Traumatology & Orthopedics, Shanghai Academy of Traditional Chinese Medicine, Shanghai 201203, China
  • Received:2023-05-23 Accepted:2023-07-17 Online:2024-09-18 Published:2023-09-28
  • Contact: Zhan Hongsheng, Professor, Chief physician, Doctoral supervisor, Shi’s Center of Orthopedics and Traumatology, Shuguang Hospital Affiliated to Shanghai University of Traditional Chinese Medicine, Shanghai 201203, China; Institute of Traumatology & Orthopedics, Shanghai Academy of Traditional Chinese Medicine, Shanghai 201203, China
  • About author:Yang Zongrui, Master candidate, Shi’s Center of Orthopedics and Traumatology, Shuguang Hospital Affiliated to Shanghai University of Traditional Chinese Medicine, Shanghai 201203, China; Institute of Traumatology & Orthopedics, Shanghai Academy of Traditional Chinese Medicine, Shanghai 201203, China
  • Supported by:
    National Natural Science Foundation of China, No. 82074466 (to ZHS); Zhan Hongsheng National Famous Elderly Chinese Medicine Experts Inheritance Workshop Construction Project, No. [2022]75 (to ZHS); Shanghai Clinical Research Center of Chronic Musculoskeletal Disease, No. 20MC1920600 (to ZHS); Shanghai Clinical Key Specialty “Chinese Medicine and Orthopedics”, No. shslczdzk03901 (to ZHS); Shanghai High-level Local Universities Innovation Team for "Chronic Muscle and Bone Disease Impairment Research and Transformation, No. [2022]3 (to ZHS)

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摘要：


文题释义：

“筋出槽”：指因慢性劳损或外力损伤，导致局部气血瘀滞，脉络瘀阻，筋脉失于濡养，筋失柔，离常道，而发为此病。最新版《ICD-11》将其明确定义：因间接暴力或慢性积累性外力作用下引起筋的形态结构、功能状态和位置关系发生异常所致；临床以患处局部疼痛、活动不利，触诊发现筋的张力增高，可触及结节、条索样改变，伴见明显压痛等为特征的伤筋病。
中医证候动物模型：基于现代实验动物学理论，将中医证候形成的病因施加于实验动物，使其模拟出具有某一证候临床特征的动物模型，从而用于中医证候相关研究。


背景：“筋出槽”是被《国际疾病分类第十一次修订本》收录的疾病名称，也是手法、针刺等治疗的临床指征，但其具体的效应机制尚不明确，亟待建立一种可以反映其临床特征和病理特点的动物模型，以便深入研究其临床治疗方法的作用机制。

目的：基于骨骼肌等长收缩建立大鼠“筋出槽”疾病动物模型，探究“筋出槽”后大鼠骨骼肌形态学和功能的特征性变化。
方法：60只大鼠随机分为对照组、静态负荷组和额外负重组，每组20只。对照组大鼠正常饲养，不做处理；后两组采用作者课题组设计的静态负荷悬挂造模装置固定大鼠，通过施加静态负荷(以大鼠自身体质量作为静态负荷)使大鼠上肢肌肉持续的等长收缩，建立“筋出槽”损伤模型；额外负重组在静态负荷组的基础上于大鼠踝关节处额外施加体质量50%的质量。分别于造模2，4周后取材，测试大鼠四肢抓力、骨骼肌湿质量、血清肌肌酸激酶和乳酸脱氢酶A水平的改变情况，观察大鼠骨骼肌组织形态学和超微结构的变化。

结果与结论：①2周时，模型组(静态负荷组和额外负重组)大鼠抓力显著降低，骨骼肌湿质量显著下降，血清肌肌酸激酶和乳酸脱氢酶A水平显著上升，肌纤维形态结构异常，伴有大量胶原纤维沉积，电镜结果显示肌原纤维结构紊乱，Z线扭曲，明暗界限模糊；②4周时，模型组大鼠抓力较2周有所上升，血清肌肌酸激酶和乳酸脱氢酶A水平下降，肌纤维形态和超微结构的改变均有不同程度恢复；③提示基于骨骼肌持续等长收缩建立的大鼠骨骼肌损伤模型，2周时可良好反映“筋出槽”的病理特点，可用于进行针刺、手法等治疗“筋出槽”的效应机制研究。

https://orcid.org/0009-0002-5643-8311(杨宗睿)

中国组织工程研究杂志出版内容重点：组织构建；骨细胞；软骨细胞；细胞培养；成纤维细胞；血管内皮细胞；骨质疏松；组织工程

关键词: “筋出槽”, 中医证候, 骨骼肌, 等长收缩, 肌酸激酶, 乳酸脱氢酶, 动物模型

Abstract: BACKGROUND: “Tendon off-position” is a disease name included in the International Classification of Diseases 11th Revision, and also a clinical indication of manipulation, acupuncture and other treatments. However, its specific mechanism is still unclear. It is urgent to establish an animal model that can reflect the clinical and pathological characteristics of “tendon off-position,” so as to further study the mechanism of effective clinical treatments.
OBJECTIVE: To establish an animal model of “tendon off-position” in rats based on isometric contraction of skeletal muscles, and to explore the changes of skeletal muscle function and morphological phenotype after “tendon off-position.”
METHODS: Sixty rats were randomly divided into control group, static-loading group and extra loading group, with twenty rats in each group. Rats in the control group were kept normally without treatment. In the latter two groups, the rats were fixed by the self-made static-loading modeling device and a static-loading (the body mass of each rats was applied as the static-loading) was applied to cause sustained isometric contraction of the upper limb muscles. Then, animal models of “tendon off-position” were successfully established. In the extra loading group, 50% of the body mass was added to the ankle joint after modeling. The skeletal muscle samples were harvested at 2 and 4 weeks after modeling. The changes of limb grip strength, wet mass of skeletal muscle, and serum levels of creatine kinase-muscle and lactate dehydrogenase A were measured, and the changes of skeletal muscle histomorphology and ultrastructure were observed.
RESULTS AND CONCLUSION: At 2 weeks after modeling, the rats in the static-loading group and extra loading group showed significantly decreased grip strength and wet muscle mass, significantly increased serum levels of creatine kinase-muscle and lactate dehydrogenase A, and abnormal muscle fiber morphology and structure accompanied by a large number of deposited collagen fibers. Electron microscopy results showed that the structure of myofibrils was disordered, the Z-line was distorted, and the light and dark boundaries were blurred. At 4 weeks after modeling, the grip strength of the model rats was increased compared with that at 2 weeks, the serum creatine kinase-muscle and lactate dehydrogenase A levels were decreased, and the changes of muscle fiber morphology and ultrastructure were recovered to varying degrees. It is suggested that the rat skeletal muscle injury model based on continuous isometric contraction of skeletal muscle can well reflect the pathological characteristics of “tendon off-position” at 2 weeks, and can be used to study the mechanism of acupuncture and manipulation in the treatment of “tendon off-position.”

Key words: “tendon off-position”, TCM syndrome, skeletal muscle, isometric contraction, creatine kinase, lactate dehydrogenase, animal model

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