TGF-β/TAK1信号通路在冲击波对粘连性肩关节囊炎肌成纤维细胞转化的作用及机制研究

Adhesive Capsulitis (AC) is characterized by pain and activity limitation of joint range of motion, which leads to obvious dysfunction. The histological examination of AC indicated that the fibroblasts change into smooth muscle phenotype (myofibroblasts). The advancement of the condition seems to be due to the presence of adhesion-related cytokines, that is, transforming growth factor β (TGF-β) and the TAK1 signal pathway. These cytokines are the key factors in the pathway of "neuropeptide—mast cell—fibroblast—myofibroblast" in the mechanism of connective tissue fibrosis. Extracorporeal Shock wave has been widely used in musculoskeletal rehabilitation, and our latest clinical study shows that shock wave can improve the pain and the activity of AC patients. The mechanism is whether the shock wave broke the continuous process of the connective tissue fibrosis, by the effect of the signal pathway of TGF-β/TAK1，thereby reducing the transformation from the fibroblasts to the myofibroblasts, there is still not clear. Therefore, in the current study, after the AC animal models preparation, the effect of TGF-β/TAK1 signal on the myofibroblast differentiation and the metabolism of ECM in vitro will be detected; and by application of shock wave on AC animal models, the myofibroblast differentiation and the expression of TGF-β1, TAK1 will be detected, to explore the mechanism of the effect of shock wave on myofibroblast differentiation of AC by TGF-β/TAK1 signal pathway,. It will provide a reliable theoretical basis for clinical shock wave treatment.

粘连性肩关节囊炎（AC），表现为活动受限及疼痛。AC的组织学检查提示成纤维细胞转变成平滑肌表型(肌成纤维细胞)，这种转变似乎与转化生长因子β(TGF-β)及TAK1信号通路的作用相关，而这些因子是结缔组织纤维粘连发生机制中“神经肽--肥大细胞--成纤维细胞--肌成纤维细胞”作用轴中的关键因子。冲击波现已广泛应用于肌骨康复中，我们的临床研究结果表明：冲击波可改善AC患者的疼痛及活动度，我们设想其作用机制可能是冲击波作用后通过抑制TGF-β/TAK1信号通路减少或逆转肌成纤维细胞的转化，打破纤维化的持续进程，影响基质胶原的代谢，重建平衡。因此，本项目拟通过AC动物模型，通过体外试验阐明TGF-β/TAK1通路对肌成纤维细胞转化及基质胶原代谢的影响，并通过体内试验探索冲击波是否通过TGF-β/TAK1通路影响肌成纤维细胞转化及其基质胶原代谢，影响纤维化的进程，揭示其作用机理。

冲击波作用于粘连性肩关节囊炎的作用机制未明，本研究通过粘连性肩关节囊炎动物模型的建立，研究冲击波对粘连性肩关节囊炎动物模型肌成纤维细胞转化及基质胶原代谢的影响，进一步探索冲击波影响肌成纤维转化及基质胶原代谢的可能信号通路，明确冲击波对于粘连性肩关节囊炎作用的可能机制。本研究通过粘连性肩关节囊炎的动物模型，发现腋下结缔组织中α-SMA蛋白表达上调，纤维结缔组织面积占比增加，MMP-2表达下调，TIMP-1/TIMP-2表达上调，提示肌成纤维细胞转化增多，纤维沉积，结缔组织金属蛋白表达失衡；冲击波作用于粘连性肩关节囊炎后，腋下结缔组织面积纤维占比下降，TGF-β、α-SMA蛋白表达下降，TAK1表达下降，提示肌成纤维细胞转化减少，同时发现冲击波作用后粘连性肩关节囊炎结缔组织MMP-2表达上调，TIMP-1、TIMP-2表达下调，表达趋向于正常组织，Ⅰ型胶原蛋白及Ⅲ型胶原蛋白表达下调，提示冲击波作用后肌成纤维细胞转化减少，MMP-TIMP平衡改善，基质代谢增加，胶原沉积减少。因此，冲击波作用于粘连性肩关节囊炎可能通过TGF-β/TAK1信号通路减少肌成纤维细胞转化，使纤维沉积下降，同时优化促进了基质金属蛋白表达代谢的平衡，从而获得了治疗粘连性肩关节囊炎的疗效。该研究为冲击波作用于粘连性肩关节囊炎的临床应用提供基础理论依据。