这项研究探讨了长链非编码RNA(lncRNA)XIST(X-失活特异性转录本)作为RA发病机理的驱动因素的作用。特别关注这种lncRNA与GATA1和CCN6相互作用的能力。下载GSE83147和GSE181614数据集用于分析。XIST和CCN6表达在滑膜成纤维细胞(SF)和正常软骨样品和RA患者中进行评估。另外检查XIST和CCN6之间的关系。XIST和CCN6在SF中分别敲低或过表达,以在RA的背景下在这些细胞中建立它们的调节作用。进一步研究XIST之间的监管相互作用,然后通过RNA免疫沉淀进行GATA1和CCN6,RNA下拉,函数增益,失去功能,和荧光素酶报告基因测定。此外,建立RA大鼠模型,测定TNF-α的产生,IL-6和IL-8,并对来自这些动物的组织进行组织病理学检查。RA患者滑膜组织和SF表现出XIST和CCN6上调。XIST的击倒抑制了SF迁移,增殖性,侵入性,和血管生成活性,而CCN6敲低部分逆转了XIST在体外和体内影响这些表型结果的能力。在SF内绑定到GATA1的XIST,从而促进CCN6转录增强。敲除XIST可减轻RA相关病理损害,滑膜损伤,和大鼠的炎症反应诱导。XIST与GATA1的结合导致CCN6上调,通过改变SF增殖和血管生成活性来驱动RA的发病机制,这表明该途径可能是治疗干预的可行目标。
This study explored the role of the long non-coding RNA (lncRNA) XIST (X-inactive specific transcript) as a driver of RA pathogenesis, with a particular focus on the ability of this lncRNA to interact with GATA1 and
CCN6. The GSE83147and GSE181614 datasets were downloaded for analysis. XIST and CCN6 expression were assessed in synovial fibroblasts (SFs) and in both normal cartilage samples and those from RA patients, with the relationship between XIST and
CCN6 additionally being examined. XIST and CCN6 were respectively knocked down or overexpressed in SFs to establish their regulatory roles in these cells in the context of RA. Further studies of the regulatory interplay between XIST, GATA1, and
CCN6 were then performed through RNA immunoprecipitation, RNA pull-down, gain-of-function, loss-of-function, and luciferase reporter assays. In addition, RA model rats were established and used to measure the production of TNF-α, IL-6, and IL-8 and to subject tissues from these animals to histopathological examination. RA patient synovial tissues and SFs exhibited XIST and
CCN6 upregulation. The knockdown of XIST suppressed SF migratory, proliferative, invasive, and angiogenic activity, while CCN6 knockdown partially reversed the ability of XIST to influence these phenotypic outcomes in vitro and in vivo. XIST bound to GATA1 within SFs, thus promoting enhanced
CCN6 transcription. Knocking down XIST alleviated RA-related pathological damage, synovial injury, and inflammatory response induction in rats. The binding of XIST to GATA1 leads to CCN6 upregulation, driving RA pathogenesis by altering SF proliferation and angiogenic activity, suggesting that this pathway may represent a viable target for therapeutic intervention.