同种异体眼外肌卫星细胞移植治疗眼外肌机械损伤的实验研究

..Therapies of paralytic strabismus result from extraocular muscles injury were challenging in ophthalmologic clinic.Transplantation of extraocular muscle satellite cell (ESC) with adult stem cells characteristics for reconstitution of functional contractile apparatus assures the regenerative capacity of exocular muscles is a promising research directions in this area. Improving the muscle regeneration ability of the ESC graft and maintaining its self-renewal in recipients are both the cruxes of the topic. We focus on acquiring optimal hierarchical composition of differentiating progenitors and selfrenewable quiescen ESCs by cytokine and microenvironment in vitro to maintains the ESC population in graft and provides proliferate, differentiate, leading to efficient new myofiber formation.Based on our preliminary research on myogenesis regulatory mechanisms including the effect of the overexpression of MAML in Notch signaling pathway, DNA methylation inhibitor CDA-II，transcription inhibitor SFMBT1 and Six1 on cell fate determination, proliferation and differentiation,we aim to build different cell culture and amplification conditions in vitro to acquire sufficient quantity of ESCs with optimal hierarchical cell composition.The ablility of muscle regeneration and functional reconstruction of the implanted ESC in the impaired extraocular muscle would be further in evaluated in rabbits animal model of extraocular muscle mechanical damage.The experiemental research could provide valuable data and method for further study of ESC transplantation to treat extraocular muscle mechanical damage.

眼外肌损伤造成的麻痹性斜视是眼科临床的棘手问题。.通过移植具有成体干细胞功能的眼外肌卫星细胞（ESC）再生眼外肌的肌纤维来重建眼外肌的收缩功能，是麻痹性斜视治疗的重要研究方向。提高移植的ESC在受体肌组织内的成肌能力，并维持ESC的自我更新是目前该领域研究的热点。.在体外通过细胞因子和培养微环境诱导扩增ECS，获取休眠状态和不同增殖分化状态构成合理的种子细胞源，是维持ESC移植后持续成肌能力的关键。.为此，我们在前期研究Notch信号通路MAML蛋白、DNA甲基化抑制剂CDA-II、转录抑制剂SFMBT1以及Six1过度表达对肌分化和增殖调控作用的基础上，拟在体外构建不同的ESC培养扩增条件，以获取数量充分和增殖分化状态合适的ESC谱系。并进一步通过兔机械性眼外肌损伤动物模型，评价ESC移植对受体损伤眼外肌组织的再生和重建作用，为通过眼外肌卫星细胞移植治疗麻痹性斜视提供实验数据。

肌卫星细胞是肌肉组织中存在的具有分化潜能的细胞，其因在肌肉的修复、再生及肥大中发挥重要作用而备受关注。眼外肌损伤造成的麻痹性斜视是眼科临床的棘手问题，利用ESC再生眼外肌的肌纤维来恢复眼外肌的收缩功能，是麻痹性斜视治疗的潜在研究方向。本研究中，我们通过对小鼠骨骼肌卫星细胞原代培养的探索，实现在临床斜视矫正术中截除的肌肉组织当中提取原代卫星细胞，并实现体外扩增。.本研究结果显示，黄酮类药物对成肌细胞系分化成肌纤维的能力有着确切的促进作用，表明黄酮类药物可以增强受损的肌肉力量，在兔眼外肌机械损伤模型当中，运用黄酮类药物可促进肌肉卫星细胞体内分化进程，进而证明其可促进眼外肌的修复，根据这项初步研究的结果，我们认为黄酮类药物可能促进肌球蛋白的形成，这有助于优化肌动蛋白-肌球蛋白的交联和修复受损的EOM。.我们的研究提示可以通过药物注射的方式，可以改善眼外肌卫星细胞的分化情况，进而促进肌肉收缩功能的恢复。这为未来斜视治疗提供了新的思路，药物注射治疗可能会是一种更安全有效的治疗方式。