推拿扌衮法对骨骼肌钝挫伤家兔模型损伤后炎症及纤维化的影响

doi:10.13418/j.issn.1001-165x.2024.2.13

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中国临床解剖学杂志 ›› 2024, Vol. 42 ›› Issue (2) : 191-195. DOI: 10.13418/j.issn.1001-165x.2024.2.13

实验研究

推拿扌衮法对骨骼肌钝挫伤家兔模型损伤后炎症及纤维化的影响

阮磊，黄博，王兰兰，薛惠天，孙梦龙，段苗苗，彭亮*

作者信息 +

The effect of tui na rolling on post-injury inflammation and fibrosis in rabbits in a model of blunt contusion of skeletal muscles

Ruan Lei, Huang Bo, Wang Lanlan, Xue Huitian, Sun Menglong, Duan Miaomiao, Peng Liang*

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摘要

目的探讨推拿扌衮法对家兔骨骼肌钝挫伤后相关纤维蛋白表达的影响，以及对骨骼肌钝挫伤修复的作用机制。方法健康成年新西兰兔15只，随机分为空白组（A）、模型组（B）和治疗（C）组，每组5只。使用自制改良重力锤打击装置对模型组和治疗组建立骨骼肌钝挫伤模型，治疗组家兔于造模成功后7 d给予推拿扌衮法干预，频率140次/min，2次/d，3 min/次，共治疗3 d。干预结束后1 d进行取材。HE和Masson染色观察兔股四头肌病理改变；Western-blot检测各组股四头肌基质金属蛋白酶-9（matrix metalloproteinases-9，MMP-9）、特异性抑制剂-1（tissue inhibitor of metalloproteinases-1，TIMP-1）、转化生长因子-β1（transforming growth factor-β1，TGF-β1）及I型胶原蛋白（I collagen，COL-I）表达。结果空白组肌组织结构规整；模型组肌组织形态迥异，边界模糊，间隙显著增宽，炎细胞浸润显著，四周胶原纤维生成增加；治疗组肌组织结构相对完整，间隙缩小，可见少量炎细胞浸润与胶原纤维生成，其病理修复程度显著优于模型组。检测家兔股四头肌MMP-9、TIMP-1、TGF-β1、COL-I表达，与空白组相比，模型组与治疗组明显增加（P<0.01）；但治疗组显著低于模型组（P<0.01）。结论推拿扌衮法能够通过减少细胞外基质过度沉积抑制骨骼肌纤维化，其作用机制可能与下调MMP-9、TIMP-1、TGF-β1及COL-I表达量有关。

Abstract

Objective To investigate the effect of tui na rolling method on the expression of relevant fibrosis proteins after blunt contusion injury of skeletal muscle in rabbits, and to explore the mechanism of action of tui na rolling method on the repair of blunt contusion injury of skeletal muscle. Methods Fifteen healthy adult New Zealand rabbits were randomly divided into three groups of blank (A), model (B) and treatment (C), with five rabbits in each group. Blunt contusion of skeletal muscle was modelled in rabbits of groups B and C using a self-constructed modified gravity hammer percussion device. The rabbits in group C were given the intervention 7 d of tui na rolling after successful modelling, at a frequency of 140 beats/min, 2 times/d, 3 min/session, for a total of 3 d of treatment. Tissue sampling was performed 1 d after the end of the intervention. Histological changes of rabbit quadriceps femoris muscle were observed by HE and Masson staining, and MMP-9, TIMP-1, TGF-β1, and COL-I expression of quadriceps femoris muscle of rabbits in each group were detected by Western-Blot. Results The muscle tissue of the blank group was of regular structure, the muscle tissue of the model group was of very distinct shape, with blurred margins, significantly enlarged gaps, significant invasion of inflammatory cells, and increased collagen fiber production in the surrounding area, the muscle tissue of the treatment group was relatively more intact in structure, with the gaps narrowed, and a small amount of invasion of inflammatory cells and collagen fiber production could be seen, and the degree of its pathological repair was significantly better than that of the model group. Compared with the blank group, the expression of MMP-9, TIMP-1, TGF-β1, and COL-I in quadriceps femoris muscle of rabbits in the model group and the treatment group were significantly increased (P<0.01). However, the expression of MMP-9, TIMP-1, TGF-β1, and COL-I in quadriceps femoris muscle of rabbits in the treatment group was significantly lower than that in the model group (P<0.01). Conclusions Tui na rolling method is ability to inhibit skeletal muscle fibrosis by reducing the excessive deposition of extracellular matrix, and then inhibit skeletal muscle fibrosis, and its mechanism of action maybe related to the suppression of the expression of MMP-9, TIMP-1, TGF-β1, and COL-I.

导出引用

阮磊, 黄博, 王兰兰, 薛惠天, 孙梦龙, 段苗苗, 彭亮. 推拿扌衮法对骨骼肌钝挫伤家兔模型损伤后炎症及纤维化的影响[J]. 中国临床解剖学杂志. 2024, 42(2): 191-195 https://doi.org/10.13418/j.issn.1001-165x.2024.2.13

Ruan Lei, Huang Bo, Wang Lanlan, Xue Huitian, Sun Menglong, Duan Miaomiao, Peng Liang. The effect of tui na rolling on post-injury inflammation and fibrosis in rabbits in a model of blunt contusion of skeletal muscles[J]. Chinese Journal of Clinical Anatomy. 2024, 42(2): 191-195 https://doi.org/10.13418/j.issn.1001-165x.2024.2.13

中图分类号： R274.3

参考文献

[1] 吴安林, 艾珏萍, 谢秀惠, 等. 推拿对骨骼肌损伤组织机化期TGF-β1、IL-6及TNF-α的影响[J]. 湖南中医杂志, 2020, 36(2): 136-138. DOI: 10.16808/j.cnki.issn1003-7705.2020.02.058.
[2] Martins L, Gallo CC, Honda T, et al. Skeletal muscle healing by M1-like macrophages produced by transient expression of exogenous GM-CSF[J]. Stem Cell Res Ther, 2020, 11(1): 473. DOI: 10.1186/s13287-020-01992-1.
[3] Hartmann DD, Gonçalves DF, Da Rosa PC, et al. A single muscle contusion promotes an immediate alteration in mitochondrial bioenergetics response in skeletal muscle fibres with different metabolism[J]. Free Radic Res, 2020, 54(2-3): 137-149. DOI: 10.1080/10715762.2020.1723795.
[4] Greising SM, Rivera JC, Goldman SM, et al. Unwavering pathobiology of volumetric muscle loss injury[J]. Sci Rep, 2017, 7(1): 13179. DOI: 10.1038/s41598-017-13306-2.
[5] Corona BT, Rivera JC, Greising SM. Inflammatory and physiological consequences of debridement of fibrous tissue after volumetric muscle loss injury[J]. Clin Transl Sci, 2018, 11(2): 208-217. DOI: 10.1111/cts.12519.
[6] Ismaeel A, Kim JS, Kirk JS, et al. Role of transforming growth factor-β in skeletal muscle fibrosis: a review[J]. Int J Mol Sci, 2019, 20(10): 2446. DOI: 10.3390/ijms20102446.
[7] Baik JE, Park HJ, Kataru RP, et al. TGF-β1 mediates pathologic changes of secondary lymphedema by promoting fibrosis and inflammation[J]. Clin Transl Med, 2022, 12(6): e758. DOI: 10.1002/ctm2.758.
[8] 刘杏, 魏晓菡, 邓洁, 等. 白藜芦醇通过抑制TGF-β_1、COL-Ⅰ过表达治疗骨骼肌损伤的实验研究[J]. 中国中西医结合杂志, 2020, 40(9): 1075-1081. DOI: 10.7661/j.cjim.20200315.137.
[9] Cabral-Pacheco GA, Garza-Veloz I, Castruita-De la Rosa C, et al. The roles of matrix metalloproteinases and their inhibitors in human diseases[J]. Int J Mol Sci, 2020, 21(24): 9739. DOI: 10.3390/ijms21249739.
[10]Carmeli E, Moas M, Reznick AZ, et al. Matrix metalloproteinases and skeletal muscle: a brief review[J]. Muscle Nerve, 2004, 29(2): 191-197. DOI: 10.1002/mus.10529.
[11]谢娇, 邓多喜, 陈英, 等. 推拿手法对骨骼肌损伤干预研究的进展[J]. 湖南中医杂志, 2018, 34(4): 199-201. DOI: 10.16808/j.cnki.issn1003-7705.2018.04.086.
[12]薛惠天, 王兰兰, 孙梦龙, 等. 不同时间点㨰法按摩器对骨骼肌损伤家兔炎症反应的影响[J]. 国际中医中药杂志, 2023, 45(4): 433-438. DOI: 10.3760/cma.j.cn115398-20220820-00236.
[13]王兰兰, 薛惠天, 孙梦龙, 等. 一种实验动物㨰法器: CN216365872U[P]. 2022-04-26.

[14]陈羽楠, 林丹红, 陈立典. 试析五体理论与中医运动功能[J]. 中医杂志, 2018, 59(4): 276-280. DOI: 10.13288/j.11-2166/r.2018.04.002.

[15]赖宇阳, 李楠. 基于《黄帝内经》探讨骨骼肌归属范畴[J]. 北京中医药大学学报, 2021, 44(6): 491-495. DOI: 10.3969/j.issn.1006-2157. 2021.06.002.
[16]屈庆, 张宏, 张国辉. 推拿㨰法舒经通络效应机制浅析[J]. 辽宁中医杂志,2015, 42(3): 598-600. DOI: 10.3969/j.issn.0411-8421. 2021. 12.034.
[17]陈海南, 卢园, 杨舟等. 小鱼际㨰法对骨骼肌钝性损伤家兔生长分化因子8与F-肌动蛋白的影响[J]. 环球中医药, 2022, 15(2): 205-210. DOI: 10.3969/j.issn.1674-1749.2022.02.004.
[18]卢园, 陈海南, 杨舟等. 小鱼际滚法对兔骨骼肌急性钝搓伤组织机化期GDF-8/Smad2通路的影响[J]. 湖南中医药大学学报, 2021, 41(9): 1345-1349. DOI: 10.3969/j.issn.1674-070X.2021.09.006.
[19]艾珏萍, 罗婷, 吴安林, 等. 小鱼际滚法对组织机化期骨骼肌钝性损伤家兔Fibronectin-1与CTGF-1表达的影响[J]. 湖南中医药大学学报, 2020, 40(2): 204-208. DOI: 10.3969/j.issn.1674-070X.2020.02.017.
[20]薛惠天, 王兰兰, 孙梦龙, 等. 云南白药气雾剂与推拿㨰法对兔骨骼肌损伤急性期核因子-κB信号通路及炎症因子变化的影响[J]. 上海中医药杂志,2023, 57(3): 80-86. DOI: 10.16305/j.1007-1334. 2023. 2209078.
[21]王兰兰, 薛惠天, 孙梦龙, 等. 推拿㨰法对兔骨骼肌急性钝挫伤组织TNF-α及SphK1、S1PR3表达的影响[J]. 中国中医药信息杂志, 2023, 30(6): 129-134. DOI: 10.19879/j.cnki.1005-5304.202209727.
[22]邓修元, 吴志彬, 杨忠. 骨骼肌纤维化的细胞与分子机制研究进展[J]. 中国康复理论与实践, 2014, 20(2): 142-147. DOI: 10.3969/j.issn.1006-9771.2014.02.011.
[23]Li Y, Foster W, Deasy BM, et al. Transforming growth factor-beta1 induces the differentiation of myogenic cells into fibrotic cells in injured skeletal muscle: a key event in muscle fibrogenesis[J]. Am J Pathol, 2004, 164(3): 1007-1019. DOI: 10.1016/s0002-9440(10)63188-4.
[24]Nagase H, Visse R, Murphy G. Structure and function of matrix metalloproteinases and TIMPs[J]. Cardiovasc Res, 2006, 69(3): 562-573. DOI: 10.1016/j.cardiores.2005.12.002.
[25]Chiao YA, Ramirez TA, Zamilpa R, et al. Matrix metalloproteinase-9 deletion attenuates myocardial fibrosis and diastolic dysfunction in ageing mice[J]. Cardiovasc Res, 2012, 96(3): 444-455. DOI: 10.1093/cvr/cvs275.
[26] Kobayashi T, Kim H, Liu X, et al. Matrix metalloproteinase-9 activates TGF-β and stimulates fibroblast contraction of collagen gels[J]. Am J Physiol Lung Cell Mol Physiol, 2014, 306(11): L1006-L1015. DOI: 10.1152/ajplung.00015.2014.
[27] Alameddine HS, Morgan JE. Matrix metalloproteinases and tissue inhibitor of metalloproteinases in inflammation and fibrosis of skeletal muscles[J]. J Neuromuscul Dis, 2016, 3(4): 455-473. DOI: 10.3233/JND-160183.
[28] Zimowska M, Olszynski KH, Swierczynska M, et al. Decrease of MMP-9 activity improves soleus muscle regeneration[J]. Tissue Eng Part A, 2012, 18(11-12): 1183-1192. DOI: 10.1089/ten.TEA.2011.0459.