针刀对特发性脊柱侧凸椎旁肌纤维化的抑制作用研究

Paravertebral muscles of idiopathic scoliosis(IS) were significantly abnormal, The main cause of IS was the imbalance of bilateral paravertebral muscles by the fibrosis of concave muscle contracture. The acupotomy was effective for the treatment of IS by releasing the concave side of paravertebral muscles contracture, improving spinal force balance, but the mechanisms were not clear yet. The mechanical environment has a significant effect on the biological behavior of the cells, which can induce the transformation of fibroblast into myofibroblast, which produce a large number of collagen fibers. Applicant’s previous studies found acupotomy could down-regulated the expression of TGF-β1 and CollagenⅠ/Ⅲ protein, and limit collagen fibers hyperplasia in injured skeletal muscle, and there is a close relationship between TGF-β1/Smads signal pathway and collagen fibers hyperplasia, so applicant make a speculation that acupotomy contracture of the paravertebral muscles can change the local mechanical environment, inhibit mechanical sensitive cells Fibroblast transformed into myofibroblast , regulate the collagen fibers by TGF-β1/Smads signal pathway ,and finally inhibiting paravertebral muscles fibrosis in IS. This study first explore the effects of Acupotomy on paravertebral muscles fibrosis, corrective force imbalance in the paravertebral muscles of IS by use the biomechanical, morphological and image methods and Then observe the Acupotomy effect on myofibroblast and related proteins and genes of TGF- beta 1/Smads pathway in paravertebral muscles.By analyzing the results, explore the possible molecular biological mechanism of acupotomy to improve the paravertebral muscles fibrosis of IS.

椎旁肌纤维化导致的脊柱两侧肌力失衡是特发性脊柱侧凸（IS）的重要病因。针刀通过切开松解IS凹侧纤维化挛缩的椎旁肌可有效改善局部力学环境，治疗IS疗效良好。力学环境对细胞生物学行为有重大影响，可诱导成纤维细胞（Fb）转化为肌成纤维细胞（MFb），后者大量生成胶原纤维。申请者前期研究发现针刀松解可降低局部组织张力，并显著下调受损骨骼肌中的TGF-β1和胶原蛋白，抑制肌肉纤维化，而TGF-β1/Smads信号通路与胶原纤维密切相关，因此提示针刀松解可能通过降低局部张力抑制Fb转化为MFb，通过TGF-β1/Smads通路减少胶原合成，抑制IS椎旁肌纤维化。本研究拟首先采用生物力学、病理切片及影像学方法，探索针刀降低局部张力对IS椎旁肌纤维化和脊柱形态的影响；然后观察针刀松解对IS椎旁肌中MFb及TGF-β1/Smads通路相关蛋白和基因的影响，揭示针刀抑制IS椎旁肌纤维化的分子生物学机制。

椎旁肌纤维化导致的脊柱两侧肌力失衡是特发性脊柱侧凸(IS)的重要病因，但以往研究很少涉及。在本研究中我们首先通过在IS大鼠和正常大鼠椎旁肌中注射TGF-β1中和抗体，观察胶原纤维改变对椎旁肌张力的影响，探讨胶原纤维在IS发展过程中的作用，然后观察针刀降低局部张力对IS椎旁肌纤维化和脊柱形态的影响，揭示针刀抑制IS椎旁肌纤维化的相关机制。..首先将大鼠随机分为正常抗体组（1组）、正常组（2组）、模型组（3组）和模型抗体组（4组），1,4组大鼠椎旁肌中注射TGF-β1中和抗体，2,3组大鼠注射生理盐水，干预完成后，1组大鼠出现了脊柱侧凸，而4组大鼠的脊柱侧凸得到了改善。1组凸侧椎旁肌和4组凹侧椎旁肌的张力显著下降。通过相关性分析发现，在1,2组凸侧椎旁肌中COL1含量与Cobb角呈负相关；在3,4组凹型椎旁肌中COL1含量与Cobb角呈正相关，COL3与Cobb角无显著相关性。证实了椎旁肌张力与胶原纤维，尤其是COL１有密切关系，通过调节胶原纤维含量可改变椎旁肌的生物力学性能，影响脊柱形态。..然后探索针刀松解椎旁肌改善脊柱侧凸的相关作用机制。大鼠随机分为正常组、IS组、中和抗体(NA)组、针刀(A)组和A+ NA组。治疗后各干预组Cobb角明显改善，其中A+NA组改善最明显。所有干预组凹侧椎旁肌张力均降低，其中A+NA组最显著。A+NA组对P-Smad2/3和Smad4蛋白表达的下调作用最明显，A组弱于NA组。与其他组相比，A+NA组的COL1和COL3浓度下降最明显，A组的COL1显著高于NA组。A+NA组纤维化面积最少，A组有轻度肌肉损伤，纤维化面积明显高于其他干预组。针刀松解可通过调节TGF-β1-Smads信号传导通路减少凹侧椎旁肌中的胶原纤维含量，恢复IS椎旁肌的力学失衡，最终纠正脊柱畸形。但在针刀松解过程中存在剥离瘢痕组织导致继发性纤维化的风险，结合TGF-β1中和抗体可预防相关副作用，达到更好的矫正效果。如何选择适合的时机和浓度注入TGF-β1中和抗体，调控胶原纤维处以最佳含量以保障骨骼肌达到最好的修复质量值得进一步探索。