LncRNA LOC100912373调控细胞自噬促进滑膜细胞增殖的机制及黄芪总皂 苷的干预研究

Rheumatoid arthritis (RA) is an autoimmune disease with an unknown etiology, occurring in approximately 1.0% of general population. More and more studies have suggested that long non-coding RNAs (lncRNAs) could play important roles in various biological processes and be associated with the pathogenesis of different kinds of diseases including RA. Our preliminary studies found that LOC100912373 as a long non-coding RNA (LncRNA) that is highly expressed in synovial tissue, whose sequence is highly conserved with NONHSAG099605.1 (human), bioinformatics analysis and experiments suggested miRNA-17, PDK1 was its candidate target gene, which were reported to play important roles in the RA by regulating the level of autophagy. Meanwhile, astragalosides (AST) not only significantly inhibited the proliferation of fibroblast-like synovial cells, but also significantly attenuated LOC100912373, PDK1and up-regulated the expressions of miR-17. Accordingly, we speculated that the LOC100912373 could combine with miR-17 competitively, relieve inhibiting of the targeted gene(PDK1), regulating the level of cell autophagy, then promoting the proliferation of synovial cells and accelerating the RA process. Based on our preliminary studies, we plan to use AA rats and original synovial cell as the model, to explore the molecular mechanism of LOC100912373 participating in RA and the intervention effect of AST by overexpression and RNAi transgenetic. This project will provides potential intervention targets for RA and new research ideas for Chinese medicine.

类风湿关节炎（RA）是一种常见病，发病机制尚不明确。课题组前期发现LncRNA LOC100912373与人NONHSAG099605.1序列高度保守，在佐剂性关节炎（AA）大鼠滑膜组织中显著升高，生物信息学及预实验初步证实miRNA-17、PDK1是其下游靶基因，且已有文献报道miRNA-17、PDK1参与RA发病。黄芪总皂苷（AST）可抑制滑膜细胞增殖，降低LOC100912373，PDK1水平，促进miRNA-17表达。根据ceRNA理论，我们推测LOC100912373可与miR-17竞争性结合，解除其对PDK1的抑制，调控细胞自噬，促滑膜细胞过度增殖，加速RA进程。本课题拟以AA大鼠和滑膜细胞为模型，采用过表达和敲低等策略，探讨LOC100912373参与RA发病的分子机理及益气健脾中药黄芪有效成分AST的干预作用，以期为RA提供临床潜在干预靶点，为治疗RA的中药研究提供新思路。

类风湿关节炎（rheumatoid arthritis，RA）是一种以对称性多关节炎为主要临床表现的慢性自身免疫性疾病，滑膜细胞过度增殖为其主要病理学基础，发病机制尚不完全明确，临床尚缺乏有效的治疗手段。长链非编码RNA(LncRNA) LOC100912373（简称LOC100912373）是RA发病的关键LncRNA。. 黄芪总皂苷（Astragalosides，AST）是黄芪的主要有效部位群，对RA患者和佐剂性关节炎（adjuvant arthritis，AA）大鼠均具有较好的治疗作用，且可调控LOC100912373的表达水平。. 本课题以“脾虚致痹，从脾论治”理论为指导，前期工作为基础，AA原代滑膜细胞为模型，LOC100912373为主线，细胞自噬为靶点，从ceRNA角度探讨RA的发病机制及AST的干预作用。. 研究表明，LOC100912373主要在FLSs细胞质表达，LOC100912373与miR-17-5p具有直接结合能力，PDK1是miR-17-5p下游靶基因。LOC100912373可作为ceRNA竞争性结合miRNA-17-5p，上调PDK1的表达，进一步激活mTOR，调控细胞自噬水平，促进滑膜细胞的过度增殖，参与RA的病理生理过程。AST可通过调控LOC100912373/miR-17-5p/PDK1信号轴及其下游靶基因的表达，抑制滑膜细胞的过度增殖，从而对RA发挥治疗作用。. 本课题的顺利实施将有利于从LncRNA角度阐明RA的发病机制，明确AST的作用机理，为临床RA诊疗提供潜在生物标志物和干预靶点，亦为治疗RA的中药研究提供新的思路。