炎性肌损伤将导致肌组织收缩无力，但肌无力是否干扰肌纤维Ca2+浓度、Calmodulin功能、Ca2+/Calmodulin依赖性通路，并反馈调节炎性肌损伤免疫反应尚缺乏文献报道。本课题拟制备急性（Notexin肌内注射）和慢性（C-protein皮内注射）小鼠肌损伤模型。分析损伤肌组织内Ca2+浓度、功能性Calmodulin及钙结合效能的差异。RNAi沉默或慢病毒载体过表达Parvalbumin基因，以调节损伤鼠肌组织Ca2+浓度，并采用Calmodulin激酶基因突变鼠诱导肌损伤，对比分析损伤肌组织内炎性细胞渗出、自身抗原表达、细胞因子和肌纤维MHC-I表达、引流淋巴结内T细胞、APCs细胞改变。通过上述研究，明确Ca2+、Calmodulin、Ca2+/Calmodulin依赖性激酶通路是否参与肌炎免疫调节，从而探讨钙通道、钙信号分子、钙拮抗剂用于炎性肌病辅助治疗的可行性。

Is it possible that muscle weakness caused by immune reaction of myoinjury，will interfere with intracellular calcium ions and different downstream Ca2+ dependent transcriptional pathways, and return to mediate the development of the disease? We presume that Ca2+/Calmodulin-dependent signal pathway involves in immune response in damaged skeletal muscle. Our first aim is to prepare acute (Notexin injection) and chronic (C-protein injection) muscle injury models with wild C57BL/6 mice. Using those models, we will explore the differences of intracellular Ca2+ concentration, functional Calmodulin, and Ca2+ binding ability of Calmodulin, compared to normal muscle. Our second aim is to prepare myoinjury models using Parvalbumin overexpress mice prepared by lentiviral vector (with persist lower levels of intracellular Ca2+), or Parvalbumin silence mice prepared by RNAi skill (with persist high levels of intracellular Ca2+), calmodulin-dependent kinases (CaMK) knock out mice. Using those models, to explore: (i) the characteristics of inflammatory cells in injuried muscle tissue; (ii) autiantigens and TLR3 express levels; (iii) pro- and anti-inflammatory cytokines, and the immune features of MHC-I+ myofibers; (iv) the proporation changes of T cells and antigen present cells (APCs). We wish that, this work will be helpful for confirming the roles of Ca2+ and Calmodulin in immune modulation of myoinjury, as well for exploring the possibility of Ca2+ pathway signal molecules, or calcium signaling antagonists to be taken as assisstant therapy drug of myositis.

肌损伤所触发的特定的肌内炎性环境可能干扰肌细胞Ca2+/Calmodulin依赖性的下游转录通路分子水平改变，并因此影响急性肌炎的免疫反应过程。本课题采用Notexin/Cardiotoxin肌内注射，或选用PNIPAM温敏水凝胶复合重组人骨骼肌C蛋白、HSNGLPL短肽修饰聚氨酯肌内植入，制备炎性肌损伤模型；对损伤肌的检测证实炎性肌组织Calmodulin及CaMKIV水平显著上调。经体内化学干扰, 以及体外基因沉默进一步证实，CaM-dependent 转录信号通路参与介导急性骨骼肌炎症反应。骨骼肌内持续上调的CaMKIV水平，将干预并延长肌内炎症过程，从而影响损伤肌组织的修复和再生。