基于碳量子点示踪分选高侵蚀性FLS体系解析藏红花素对侵蚀相关转录因子-miRNA前馈环路的影响

Fibroblast like synoviocytes (FLS) shows "tumor like" erosion behavior on extracellular matrix and cartilage in non-inflammatory and inflammatory environment，and is currently thought to be a primary cell mediating rheumatoid arthritis (RA) joint destruction. While, how to sort such highly invasive cells and explore the mechanism of transcriptional regulation of phenotypic stability of erosion remain serious problem. Saffron can be used to treat rheumatoid arthritis in pill, medicated bath, and patch application forms. Crocin, as the active ingredient of saffron, was found to have the ability to inhibit the invasion of FLS by the applicant for the first time. Based on fluorescence tracing and charge loading characteristics, new carbon quantum dots screening method was applied to get highly active erosion phenotype of primary FLS; the bioinformatics high-through analysis method was deployed to reveal the mechanism regulating transcription stability of FLS erosion---transcription factor-miRNA feed-forward loop; the effects of crocin on FLS and rheumatoid arthritis were observed and the change of erosion transcription factor-miRNA feed-forward loop was inferred and verified. This project will further enrich the theoretical system of saffron treatment and provide a theoretical basis for clinical application.

成纤维样滑膜细胞（Fibroblast Like Synoviocytes, FLS）在炎性和非炎性环境中均具有“肿瘤样的”侵蚀细胞外基质和软骨的能力，目前被认为是介导类风湿性关节炎(Rheumatoid Arthritis，RA)关节破坏的一种主要细胞。如何获得高侵蚀性FLS和揭示其侵蚀表型稳定的转录调控机制是亟待解决的问题。藏红花以丸剂、药浴、贴剂等形式应用于藏医RA治疗当中，申请人首次发现其活性成分藏红花素可以抑制FLS侵袭。本研究基于碳量子点荧光示踪和电荷负载特性，构建新型高侵蚀性FLS筛选体系；应用生物信息高通量分析方法揭示与FLS侵蚀相关的稳定转录机制---转录因子-miRNA前馈环路；观察体内外模型中藏红花素的作用效果及其与前馈环路间的关系，推断及验证其可能转录调控点。丰富藏红花治疗体系，为临床应用藏红花素提供理论依据。

成纤维样滑膜细胞（Fibroblast Like Synoviocytes, FLS）被认为是介导类风湿性关节炎(Rheumatoid Arthritis，RA)关节破坏的一种主要侵蚀性细胞。申请人首次发现藏红花素可以抑制FLS侵袭。本研究基于碳量子点荧光示踪和电荷负载特性，构建新型高侵蚀性FLS筛选体系；应用互联网类风湿性关节炎和骨性关节炎差异表达数据(GSE72564)和申请者构建的高低侵袭性FLS表达谱数据，对数据进一步分析发现，NFKB作为主要转录因子参与侵袭过程；应用迁移、Transwell侵袭等实验证实藏红花素可以抑制NFKB介导的FLS侵袭过程。本研究丰富了藏红花治疗体系，为临床应用藏红花素提供理论依据和实验支持。