炎症小体放大环路促进玫瑰痤疮发病的效应与机制研究

Rosacea is a kind of refractory skin disease which has serious influence in people’s appearance and healthy. It is of paramount important to find that innate immune response is over-activated in rosacea. Recent research reveals that the excreted ASC-Speck could be internalized by bystander cells, which leads to NLRP3 activation and the auto-amplification mechanism of inflammasome. In our previous clinical study, it was found that the severity of inflammatory papulopustular rosacea was negatively related with the numbers of demodex folliculorum, while positvely related with ASC-Speck status. It was aslo demonstrated that in vitro experiments, the macrophage dirved ASC-Speck and then internalized by fibroblast 3T3 cells, which directly activates inflammasome and IL-1β production. Base these findings, our research team propose a hypothesis that there is an inflammation amplification system, which transduces epidermal microorganism infection into dermal inflammation. Said another way, demodex folliculorum induced first inflammation response in epidermis, which could be transduced into dermis by internalization of ASC-Speck. In consequence, our research team are planning to investigate the NLRP3-medicated inflammasome and its amplification system for rosacea’s development. Four levels will be proceeded in this research: clinical assessments, mice rosacea model testing, cell function in vitro and signal transduction assay. Our research will provide theoretical basis for rosacea treatment and supply the novel candidate for rosacea’s medicine design.

玫瑰痤疮是严重影响人群美观的顽固性皮肤疾病。天然免疫反应过度活化是其重要病因。新近研究发现NLRP3介导炎症小体回收ASC聚合体实现自我放大环路，促进炎症发展。我们前期研究发现：丘疹脓疱型玫瑰痤疮患者的局部炎症反应程度与毛囊蠕形螨数量负相关，与ASC聚合体程度正相关；体外细胞实验发现成纤维细胞株3T3细胞内吞ASC-Speck小体直接活化炎症小体。基于以上结果课题组假设：皮肤浅层微生物感染与深层炎症反应之间存在一放大系统的连接，即皮肤中毛囊蠕形螨等病原体刺激毛囊皮脂腺相关细胞产生第一步炎症反应，真皮层成纤维细胞可回收ASC-Speck小体直接活化炎症小体，产生第二步炎症反应，导致真皮炎症反应。课题组拟通过临床样本、玫瑰痤疮小鼠模型、体外细胞与信号转导分析，探索玫瑰痤疮的发展中，NLRP3介导的炎症小体及其放大环路中的作用与分子机制，为玫瑰痤疮发病机制提供理论基础，也为开发药物提供设计靶点。

玫瑰痤疮发病机制一直是皮肤科关注重点。而作为天然免疫重要环节之一的炎症小体放大环路，是否和玫瑰痤疮发生发展具有重要相关性，存在很多研究价值。.本研究从NLRP3炎症小体活化程度、ASC-Speck聚合体与皮肤炎症反应强弱相关性入手，通过动物模型确定炎症小体活化参与了玫瑰痤疮的发生发展，并结合细胞相关实验，来证实ASC-Speck介导的炎症小体放大环路可一定程度上促进玫瑰痤疮动物模型的发展。.重要结果和关键数据：.1.临床上，完善皮肤共聚焦显微镜评价技术，对玫瑰痤疮和其他相关疾病患者进行反复多次检测，寻找出皮肤共聚焦显微镜下最合适的评价参数：如表皮角化异常、蜂窝样结构、海绵状水肿和真皮乳头改变是疾病确诊的重要依据，并且对于疾病的严重程度有重要的参考价值；.2.完成了玫瑰痤疮动物模型制备：对模型制备进行相关具体方法学的调整，如具体的给药注射剂量、使用的方法、观察时间等，在实验组中出现明显的玫瑰痤疮样红斑，并且病理结果中证实有炎症性改变；完成免疫组化相关检测结果，并且有了相关定量数据，LL37+PBS、LL37+ASC-speck、LL-37+VX-765之间具有显著性差异；NLRP3炎症小体水平、MMP9水平以及 Caspase 1水平，在不同的模型中均有显著性差异；.3.细胞实验功能与信号转导研究相关工作已经完成，数据也在进一步整理中。.科学意义：.本研究从临床评价、到动物模型建立、到细胞实验功能，对于了解玫瑰痤疮的发展过程中，微生物感染刺激毛囊皮脂腺相关细胞产生第一步炎症反应，真皮层成纤维细胞回收ASC-Speck小体直接活化炎症小体产生第二步炎症反应。炎症小体放大环路是玫瑰痤疮发展过程中的炎症放大的可能重要机制之一。这对于深入理解玫瑰痤疮发病机制，寻求新的治疗方案有了更多实验室依据。.目前已经发表SCI文章5篇，其中3篇有此基金标注；另外2篇相关文章正在投稿中。