紫草经线粒体AMPK/ROS通路调控NFAT5改善糖尿病难愈性创面“热气留滞”的作用机制

炎症持续是糖尿病创面难愈的主要原因，紫草油清热敛疮，减轻炎症促创面愈合，热气留滞是病机关键。糖尿病是皮肤高渗主要的内在病因，创面发生与高渗密切相关。活化T细胞核因子5（NFAT5）是在高渗下启动并放大炎症的转录因子，高表达于糖尿病致创面难愈。前期研究发现NFAT5在糖尿病早期皮肤及创面高表达，紫草抑制NFAT5。结合线粒体AMPK/ROS通路在调控NFAT5促炎功能中重要作用，我们提出：紫草可经AMPK/ROS选择性抑制NFAT5炎症转录，减少炎症因子释放，使NFAT5恢复生理功能，改善热气留滞促进创面愈合。项目以角质形成细胞、糖尿病鼠创面模型为对象，通过免疫组化等技术，设抑制或过表达组，检测AMPK/ROS/NFAT5通路信号分子变化，揭示紫草油清热敛疮的微观机制。本项目以NFAT5炎症调控热气留滞的新视点揭示紫草促糖尿病创面愈合的机制，异病同治，为紫草在其他高渗相关皮肤病应用提供依据。

Persistent inflammation is the main cause of impaired healing of diabetic wounds. Zicao oil clears heat and collects sores, relieves inflammation and promotes wound healing. Heat retention is the key to pathogenesis. Diabetes mellitus is the main internal cause of skin hyperosmosis, and wound occurrence is closely related to hyperosmosis. Nuclear factor of activated T cells 5 (NFAT5) is a transcription factor that can initiate and amplify inflammation under hyperosmotic stress and is highly expressed in diabetes mellitus，leading to wounds hard healing. Preliminary experiments showed that NFAT5 was highly expressed in early diabetic skin and wounds, and Zicao could inhibit the expression of NFAT5 mRNA and protein. As mitochondrial AMPK/ROS/NFAT5 pathway plays an important role in regulating the pro-inflammatory function of NFAT5, we hypothesized that, through AMPK/ROS/NFAT5 pathway, Zicao could selectively inhibit the transcription region of NFAT5 inflammatory factors, reduce the release of inflammatory factors, alleviate inflammation, restore NFAT5 physiological function, and promote diabetic wound epithelialization. Keratinocytes and diabetic mouse wound models will be taken as research object. Through immunohistochemistry and other techniques, the inhibition or overexpression group will be set, the effects on expression of AMPK/ROS/NFAT5 signaling molecules in diabetic wounds will be studied. The expression changes of AMPK/ROS/NFAT5 signaling molecules in diabetic wounds under different conditions will be detected, so that reveal the mechanism of heat-clearing and sores fetching effect of Zicao oil. This project reveals a novel mechanism of Zicao in treating refractory diabetic wounds from the new perspective of NFAT5 inflammation regulation, and clarifies the relationship between Zicao and hyperosmotic stress mechanism, treating different diseases with same method, which can provide theoretical basis for the application of Zicao in other hyperosmotic-related skin diseases.