miR-21/PDCD4/NF-κB通路在血小板抗菌促糖尿病溃疡愈合中的作用及分子机制

项目名称： miR-21/PDCD4/NF-κB通路在血小板抗菌促糖尿病溃疡愈合中的作用及分子机制

项目编号： No.81500596

项目类型： 青年科学基金项目

立项/批准年度： 2016

项目学科： 医药、卫生

项目作者： 邓武权

作者单位： 中国人民解放军第三军医大学

项目金额： 18万元

中文摘要： 感染是阻碍糖尿病溃疡（DU）愈合的关键因素。研究提示富血小板血浆 (PRP) 具有抗感染促DU愈合作用，但其机制尚不清楚。新近研究提示miR-21参与调控NF-κB发挥抗感染促修复作用。我们前期研究发现小鼠金黄色葡萄球菌（S. aureus）DU愈合中miR-21表达下降；PRP干预后miR-21、IL-10上调，而PDCD4和NF-κB下调。结合文献调研，我们提出PRP通过miR-21调控PDCD4和NF-κB的表达及通路活化发挥拮抗感染性炎症并促进DU愈合的假设。本项目拟以HaCaT细胞和小鼠S. aureus DU为模型，选用Northern blot等方法离体研究miR-21与PDCD4的相互作用及对下游分子表达和NF-κB信号通路活化的影响；在体验证miR-21抗感染促愈合作用。期望揭示血小板分泌miR-21在PRP治疗DU中的作用及分子机制，为防治难愈性创面提供依据和新靶点。

中文关键词： 小分子RNA-21；血小板；糖尿病溃疡；创面感染；信号通路

英文摘要： Infection is one of the contributing factors for non-healing diabetic ulcer (DU) . Thus exploring new antimicrobial strategy poised an important practical significance. Studies suggested that platelet-rich plasma (PRP) can promote DU healing with anti-inflammatory function, but its mechanism is unclear. Recent studies revealed that miR-21 plays a key effect on antimicrobial, cell proliferation and migration via regulation of nuclear transcription factor (NF-κB) expression. Our previous study found that miR-21 expression was down-regulated in Staphylococcus aureus (S. aureus) infested mice during wound healing process. After PRP intervention, only miR-21 and IL-10 expression were up-regulated, while PDCD4 or NF-κB still down-regulated. Extrapolating from these preliminary experiment results, we propose a hypothesis: PRP promotes DU anti-microbial and wound healing through regulating PDCD4 expression and NF-κB pathway activation. The study intends to observe miR-21 regulates PDCD, NF-κB expression and IL-6, IL-10, TGF-β1 secretion through different glucose levels of HaCaT cells and infected mice DU induced by S. aureus. Further, miR-21 interacts with PDCD4 and its effect on downstream molecules expression and NF-κB signaling pathway activation through Northern blot and RT-PCR in vitro study. Meanwhile, the influence of miR-21 on DU antimicrobial and wound healing will be investigated in vivo study. Through above research, it is expected to reveal molecular mechanisms and healing effects of miR-21 released by platelet for DU treatment, which may provide a theoretical basis and new targets for prevention and treatment of recalcitrant wound.

英文关键词： miRNA-21;platelet;diabetic ulcer ;wound infection;signaling pathway