中国组织工程研究 ›› 2025, Vol. 29 ›› Issue (8): 1600-1608.doi: 10.12307/2025.337

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

理筋手法调控兔骨骼肌损伤修复中瘢痕形成的作用机制

李开颖1,2,3,魏晓歌1,2,3,宋  斐1,2,3,杨  楠1,2,3,赵振宁1,2,3,王  燕1,穆  静1,马惠昇1,2,3   

  1. 宁夏医科大学,1中医学院,2宁夏少数民族医药现代化教育部重点实验室,宁夏回族自治区银川市  750004;3宁夏区域高发病中西医结合防治研究重点实验室,宁夏回族自治区银川市  750004
  • 收稿日期:2024-01-12 接受日期:2024-04-09 出版日期:2025-03-18 发布日期:2024-07-05
  • 通讯作者: 马惠昇,博士,教授,宁夏医科大学,中医学院,宁夏少数民族医药现代化教育部重点实验室,宁夏回族自治区银川市 750004; 宁夏区域高发病中西医结合防治研究重点实验室,宁夏回族自治区银川市 750004 并列通讯作者:穆静,博士,副教授,宁夏医科大学中医学院,宁夏回族自治区银川市 750004
  • 作者简介:李开颖,女,1991年生,宁夏回族自治区银川市人,回族,宁夏医科大学在读硕士,主要从事筋伤疾病基础与临床研究。
  • 基金资助:
    国家自然科学基金(82160944),项目名称:基于Wnt信号通路研究理筋手法对骨骼肌损伤的神经肌肉接头功能重塑机制,项目负责人:马惠昇

Mechanism of Lijin manipulation regulating scar formation in skeletal muscle injury repair in rabbits

Li Kaiying1, 2, 3, Wei Xiaoge1, 2, 3, Song Fei1, 2, 3, Yang Nan1, 2, 3, Zhao Zhenning1, 2, 3, Wang Yan1, Mu Jing1, Ma Huisheng1, 2, 3   

  1. 1School of Traditional Chinese Medicine, Ningxia Medical University, Yinchuan 750004, Ningxia Hui Autonomous Region, China; 2Ningxia Medical University Key Laboratory of Ningxia Minority Medicine Modernization Ministry of Education, Yinchuan 750004, Ningxia Hui Autonomous Region, China; 3Ningxia Regional Key Laboratory of Integrated Traditional Chinese and Western Medicine for Prevention and Treatment of Regional High Incidence Disease, Yinchuan 750004, Ningxia Hui Autonomous Region, China
  • Received:2024-01-12 Accepted:2024-04-09 Online:2025-03-18 Published:2024-07-05
  • Contact: Ma Huisheng, MD, Professor, School of Traditional Chinese Medicine, Ningxia Medical University, Yinchuan 750004, Ningxia Hui Autonomous Region, China; Ningxia Medical University Key Laboratory of Ningxia Minority Medicine Modernization Ministry of Education, Yinchuan 750004, Ningxia Hui Autonomous Region, China; Ningxia Regional Key Laboratory of Integrated Traditional Chinese and Western Medicine for Prevention and Treatment of Regional High Incidence Disease, Yinchuan 750004, Ningxia Hui Autonomous Region, China Co-corresponding author: Mu Jing, MD, Associate professor, School of Traditional Chinese Medicine, Ningxia Medical University, Yinchuan 750004, Ningxia Hui Autonomous Region, China
  • About author:Li Kaiying, Master candidate, School of Traditional Chinese Medicine, Ningxia Medical University, Yinchuan 750004, Ningxia Hui Autonomous Region, China; Ningxia Medical University Key Laboratory of Ningxia Minority Medicine Modernization Ministry of Education, Yinchuan 750004, Ningxia Hui Autonomous Region, China; Ningxia Regional Key Laboratory of Integrated Traditional Chinese and Western Medicine for Prevention and Treatment of Regional High Incidence Disease, Yinchuan 750004, Ningxia Hui Autonomous Region, China
  • Supported by:
    National Natural Science Foundation of China, No. 82160944 (to MHS) 

摘要:


文题释义:
理筋手法:是以筋伤理论为依据,对筋伤部位进行针对性理筋推拿手法操作,达到舒筋活络、理筋止痛效果,从而治疗筋伤疾病的一种治疗手法。
骨骼肌损伤:在剧烈运动时造成肌肉拉伤或者挫伤,导致肌肉肿胀、疼痛、紧张、痉挛、僵硬、硬结,甚至会出现皮下淤青、按压时疼痛明显加重、损伤肌肉活动受限、拉伸时疼痛加剧等症状。属于软组织损伤,中医称之为“筋伤”。

背景:理筋手法能够促进骨骼肌修复,治疗骨骼肌损伤。但骨骼肌损伤在修复过程中的纤维化形成、瘢痕组织增生等与损伤修复质量密切相关。开展理筋手法对纤维化形成、瘢痕组织增生的调控作用研究,有利于阐述理筋手法提高骨骼肌损伤修复质量的相关机制。
目的:探索理筋手法提高兔骨骼肌损伤后修复质量的作用机制,为临床治疗提供科学依据。
方法:45只健康成年日本大耳白兔随机分为空白组、模型组、理筋组,每组15只。其中模型组和理筋组均进行腓肠肌打击造模;造模后理筋组于第3天开始进行理筋手法干预,1次/d,15 min/次。各组在造模后的第7,14,21天分别处死5只兔进行观察。苏木精-伊红染色法观察腓肠肌形态及炎性细胞量,Masson染色法观察腓肠肌胶原纤维量,ELISA法检测腓肠肌白细胞介素6和白细胞介素10的表达量,Western blot和RT-PCR检测配对盒基因7、成肌分化因子、肌细胞生成素、肌动蛋白α、转化生长因子β1、Ⅰ型胶原蛋白的蛋白及mRNA表达,免疫组织化学法检测Ⅰ型胶原蛋白的表达。
结果与结论:①苏木精-伊红染色及Masson染色结果显示,与模型组比较,理筋组各观察点炎性细胞浸润减少,胶原纤维量减少(P < 0.01),肌纤维逐渐愈合;②ELISA结果显示,与模型组比较,理筋组白细胞介素6表达持续降低(P < 0.05),而白细胞介素10在造模后第7天时升高(P < 0.05),随后呈下降趋势(P < 0.05);③Western blot和RT-PCR结果显示,与模型组比较,理筋组造模后第14天配对盒基因7、成肌分化因子、肌细胞生成素的蛋白及mRNA表达量均显著升高(P < 0.05),而第21天时却较之前下降(P < 0.05);理筋组各观察点肌动蛋白α、转化生长因子β1、Ⅰ型胶原蛋白的蛋白及mRNA表达量相较于模型组均显著降低(P < 0.05);④免疫组化结果显示,理筋组各观察点Ⅰ型胶原蛋白的表达量相较于模型组均显著降低(P < 0.05);⑤结果表明,理筋手法能够通过抑制炎症、促进肌卫星细胞的增殖分化、减少纤维化的生成,从而提高兔骨骼肌损伤的修复质量。
https://orcid.org/0009-0005-6835-562X(李开颖)

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: 理筋手法, 筋伤, 骨骼肌损伤, 炎症, 肌卫星细胞, 纤维化, 瘢痕组织, 修复质量

Abstract: BACKGROUND: Lijin manipulation can promote skeletal muscle repair and treat skeletal muscle injury. However, the formation of fibrosis and scar tissue hyperplasia are closely related to the quality of skeletal muscle repair. To study the regulatory effect of Lijin manipulation on the formation of fibrosis and scar tissue hyperplasia is helpful to explain the related mechanism of Lijin manipulation to improve the repair quality of skeletal muscle injury.
OBJECTIVE: To explore the mechanism of Lijin manipulation to improve the repair quality of skeletal muscle injury in rabbits, thereby providing a scientific basis for clinical treatment.
METHODS: Forty-five healthy adult Japanese large-ear white rabbits were randomly divided into blank group, model group and Lijin group, with 15 rats in each group. Gastrocnemius strike modeling was performed in both model group and Lijin group. The Lijin group began to intervene with tendon manipulation on the 3rd day after modeling, once a day, and 15 minutes at a time. Five animals in each group were killed on the 7th, 14th and 21st days after modeling. The morphology and inflammatory cell count of gastrocnemius were observed by hematoxylin-eosin staining, the collagen fiber amount was observed by Masson staining, the expression of interleukin-6 and interleukin-10 in gastrocnemius was detected by ELISA. The protein and mRNA expressions of paired cassette gene 7, myogenic differentiation factor, myoblastogenin, alpha-actin, transforming growth factor beta 1, and type I collagen were detected by western blot and RT-PCR, respectively, and the expression of type I collagen protein was detected by immunohistochemistry.
RESULTS AND CONCLUSION: Hematoxylin-eosin staining and Masson staining showed that compared with the model group, inflammatory cell infiltration and collagen fiber content decreased in the Lijin group (P < 0.01), and the muscle fibers gradually healed. ELISA results showed that compared with the model group, the expression of interleukin-6 in the Lijin group continued to decrease (P < 0.05), and the expression of interleukin-10 increased on the 7th day after modeling (P < 0.05) and then showed a decreasing trend (P < 0.05). Western blot and RT-PCR results showed that compared with the model group, the protein and mRNA expressions of paired cassette gene 7, myogenic differentiation factor, myoblastogenin in the Lijin group were significantly increased on the 14th day after modeling (P < 0.05), but decreased on the 21st day (P < 0.05); the protein and mRNA expressions of alpha-actin, transforming growth factor beta 1, and type I collagen in the Lijin group were significantly decreased compared with those in the model group (P < 0.05). Immunohistochemical results showed that the expression of type I collagen in the Lijin group was significantly lower than that in the model group (P < 0.05). To conclude, Lijin manipulation could improve the repair quality of skeletal muscle injury by inhibiting inflammation, promoting the proliferation and differentiation of muscle satellite cells, and reducing fibrosis.

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

Key words: Lijin manipulation, injury of tendons, skeletal muscle injury, inflammation, muscle satellite cell, fibrosis, scar tissue, repair quality


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