外层负载ERK2-siRNA的双向组织匹配性纳米纤维膜抑制肌腱粘连组织增生的相关机制研究

Peritendinous adhesion limits limb function greatly. At the process of treatment, anti-adhesion membranes can act as a physical barrier but can not promote tendon healing and sliding as well as adhesion tissue formation. Our previous study confirmed that bi-layer sheath membrane can promote tendon healing and sliding, and also confirmed that extracellular signal-regulated kinases (ERK2) is a key target of tissue adhesion formation and Polyplex can delivery small interfering RNA(siRNA) effectively. This project intends to prepare poly(l-lactic acid)- polyethylene glycol(PELA) bi-layer sheath membrane whose outer layer was loaded with ERK2-siRNA/Polyplexs by electrospinning, detect the drug release and gene interfering efficiency and reveal the effect of membranes on the fibroblast and its mechanism that whether through inhibiting ERK2 and linker region of Smad2 and Smad3 phosphorylation. Furthermore, the effect of prevention of adhesion tissue formation and its mechanism and the result of tendon healing will be investigated in a leghorn chicken model to provide a new treatment for rehabilitation of limb function and theoretical basis.

肌腱粘连严重影响患者手部功能。治疗中，既往仅有阻隔作用的防粘连膜不能促进肌腱愈合和滑动，也无法抑制组织增生导致的粘连。我们前期构建了促进肌腱愈合和滑动的双向组织匹配性防粘连膜，证实了胞外信号调节激酶2（ERK2）是抑制肌腱粘连组织增生的靶点，并合成了高效输送小干扰RNA的聚阳离子纳米颗粒（Polyplex）。在此基础之上，本课题拟采用静电纺丝技术以聚乳酸聚乙二醇共聚物为材料制成外层负载ERK2-siRNA/Polyplex的双向组织匹配性防粘连膜，体外研究药物释放特性及基因干扰效率，观察该膜对成纤维细胞增殖的抑制作用，并通过检测ERK2的磷酸化、Smad2和Smad3连接处位点的磷酸化水平对其作用机制进行深入探讨；采用鸡趾屈肌腱粘连模型，体内实验验证该膜植入后对腱周粘连组织增生的影响并探索其机制，观察对肌腱滑动及内愈合的影响。本课题为防止肌腱粘连促进患肢功能康复提供新的治疗方法和理论依据。

肌腱粘连严重影响患者手部功能。治疗中，既往仅有阻隔作用的防粘连膜不能促进肌腱愈合和滑动，也无法抑制组织增生导致的粘连。本课题利用微凝胶载药及静电纺丝技术将聚阳离子PDA与ERK2-siRNA复合的纳米颗粒（ERK2-siRNA-PDA）载入纤维内部，形成缓释给药的防粘连膜；证实该膜可以抑制UMNSAH/DF-1成纤维细胞的粘附和增殖；并在Leghorn鸡趾屈肌腱粘连模型体内证实该膜具有较好的防粘连能力，而且可能通过下调ERK2减少了粘连组织III型胶原含量。同时，还进行了水凝胶微球以及载布洛芬抗炎防粘连膜的研究，为防止肌腱粘连、促进患肢功能康复提供新的治疗方法及理论依据。