钠离子通道Nax对糖尿病难愈性创面的调控及其表观遗传学机制的研究

我们前期研究发现，表皮细胞钠离子通道Nax的表达及介导的钠离子内流，是炎症反应、皮肤真皮细胞激活的关键。进一步实验发现，真皮中内皮细胞同样有Nax表达，但在高血糖环境中，其表达显著增加，伴随细胞功能紊乱，而CRAM1的表达异常及其对NF-κB 通路的激活，可能是高糖环境下Nax调控细胞功能的主要机制。基于前期研究基础，本项目拟建立体外皮肤微血管内皮细胞高糖培养模型及糖尿病大鼠慢性创面模型，采用慢病毒双相调控体系及特异性靶向抑制剂，联合ChIP_PCR、ChIP-Seq测序及荧光素酶报告等技术，探究Nax通过CRAM1介导的蛋白质精氨酸甲基化调控NF-κB信号通路，对糖尿病创面愈合的调控及其潜在作用机制。本项目的完成，不仅从表观遗传学的角度阐述高糖环境对创面愈合的影响及机制，而且为糖尿病难愈创面的预防和治疗提供可靠的理论基础和新的治疗靶点。

Our previous studies have found that the expression of sodium channel Nax in epidermal cells and the followed sodium ion inflow were the key to excessive inflammatory response and dermal cells activation. Further experiments showed that the expression of Nax in endothelial cells in the dermis was also observed, but the expression of Nax increased significantly in the hyperglycemia environment, which was accompanied by the dysfunction of cell function. The abnormal expression of CRAM1 and the followed activation of NF-κB pathway may be the main mechanism of Nax regulating cell function in the hyperglycemia environment. Based on our previous research, this project intends to establish the in vitro skin microvascular endothelial cells high sugar cultivation model and diabetic rats model of chronic wounds, exploring the underlying mechanism of Nax/CRAM1-mediated protein arginine methylation and the following NF-κB signaling pathway activation on the regulation of diabetic wound healing by using the slow virus bipolar control system and the specific targeting inhibitor, combined ChIP_PCR, ChIP - Seq sequencing and luciferase report technologies. The completion of this project not only elaborated the effect and mechanism of high glucose environment on wound healing from the perspective of epigenetics, but also provided a reliable theoretical basis and a new therapeutic target for the prevention and treatment of diabetic refractory wound healing.