环状RELL1通过调控miR-200c-3p促进骨关节炎进展。
CircRELL1 promotes osteoarthritis progression by regulating miR-200c-3p.
作者信息
Ding HongZhi, Chen HaiJu, Dou LianRong, Li Yang
机构信息
Department of Orthopedic, Shanghai Songjiang District Central Hospital, Shanghai, 201699, China.
Department of Orthopedic, Zhongshan Hospital Affiliated to Fudan University, Shanghai, 200032, China.
出版信息
Heliyon. 2024 Jul 6;10(14):e34251. doi: 10.1016/j.heliyon.2024.e34251. eCollection 2024 Jul 30.
BACKGROUND
There is a growing body of evidence indicating a potential association between circular RNA and the pathogenesis of human osteoarthritis (OA). Nevertheless, the precise extent of their involvement in OA remains largely unexplored. Hence, the objective of this investigation is to elucidate the function of Circular (Circ) RELL1 in the context of OA.
METHODS
24 OA tissue samples and 11 normal tissue samples were collected. The inflammatory OA-like conditions were established by Destabilized Medial Meniscus (DMM) operation in mice and LPS-induced C28/I2 cells. OA severity and articular cartilage degradation were assessed by Safranin-O staining, hematoxylin-eosin (H&E) staining, and International Society for Osteoarthritis Research (OARSI) criteria. CircRELL1, miR-200c-3p, and TCF4 were measured by RT-qPCR and Immunoblot. The cell viability and apoptosis rate were measured by MTT and flow cytometry, respectively. The levels of cytokines interleukin (IL)-1β, IL-6, and TNF-α were determined by ELISA. Apoptosis-associated proteins (cleaved caspase-3, Bax, and Bcl-2) and extracellular matrix (ECM) degradation-associated proteins (MMP13, collagen II, and Aggrecan) were detected by Immunoblot. The interaction between miR-200c-3p and circRELL1 or TCF4 was verified by dual luciferase reporter assay and RIP assay.
RESULTS
CircRELL1 expression was upregulated in OA patients, and the results were consistent in DMM mice and LPS-treated C28/I2 cells. Silencing circRELL1 improved cartilage injury caused by DMM and contributed to a lower OARSI score. Silencing CircRELL1 increased the activity of OA chondrocytes and and inhibited cellular inflammatory responses and ECM degradation. In terms of mechanism, circRELL1 functioned by targeting miR-200c-3p, leading to the suppression of inflammatory factor production, cell apoptosis, and ECM degradation, thus inhibiting the progression of OA.
CONCLUSION
CircRELL1 may promote the progression of OA by regulating the miR-200c-3p.
背景
越来越多的证据表明环状RNA与人类骨关节炎（OA）的发病机制之间存在潜在关联。然而，它们在OA中的具体参与程度仍 largely未被探索。因此，本研究的目的是阐明环状（Circ）RELL1在OA背景下的功能。
方法
收集24例OA组织样本和11例正常组织样本。通过在小鼠中进行内侧半月板不稳定（DMM）手术和脂多糖诱导C28/I2细胞建立炎症性OA样病症。通过番红O染色、苏木精-伊红（H&E）染色和国际骨关节炎研究学会（OARSI）标准评估OA严重程度和关节软骨降解情况。通过RT-qPCR和免疫印迹法检测CircRELL1、miR-200c-3p和TCF4。分别通过MTT法和流式细胞术检测细胞活力和凋亡率。通过ELISA法测定细胞因子白细胞介素（IL）-1β、IL-6和肿瘤坏死因子-α的水平。通过免疫印迹法检测凋亡相关蛋白（裂解的半胱天冬酶-3、Bax和Bcl-2）和细胞外基质（ECM）降解相关蛋白（MMP13、胶原蛋白II和聚集蛋白聚糖）。通过双荧光素酶报告基因测定法和RIP测定法验证miR-200c-3p与circRELL1或TCF4之间的相互作用。
结果
CircRELL1在OA患者中表达上调，在DMM小鼠和脂多糖处理的C28/I2细胞中结果一致。沉默circRELL1可改善DMM引起的软骨损伤并导致较低的OARSI评分。沉默CircRELL1可增加OA软骨细胞的活性并抑制细胞炎症反应和ECM降解。在机制方面，circRELL1通过靶向miR-200c-3p发挥作用，导致炎症因子产生、细胞凋亡和ECM降解受到抑制，从而抑制OA的进展。
结论
CircRELL1可能通过调节miR-200c-3p促进OA的进展。
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