■越来越多的证据表明环状RNA与人类骨关节炎(OA)的发病机理之间存在潜在关联。然而,他们参与OA的确切程度在很大程度上仍未被探索。因此,本研究的目的是阐明循环(Circ)RELL1在OA中的作用.
■收集24个OA组织样品和11个正常组织样品。在小鼠和LPS诱导的C28/I2细胞中,通过去稳定的内侧半月板(DMM)手术建立了炎性OA样条件。通过Safranin-O染色评估OA严重程度和关节软骨降解,苏木精-伊红(H&E)染色,和国际骨关节炎研究协会(OARSI)标准。CircRELL1,miR-200c-3p,通过RT-qPCR和免疫印迹测量TCF4。MTT法和流式细胞术检测细胞活力和凋亡率,分别。细胞因子白细胞介素(IL)-1β,通过ELISA测定IL-6和TNF-α。凋亡相关蛋白(裂解的caspase-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Aggrecan)通过免疫印迹检测。miR-200c-3p与circRELL1或TCF4之间的相互作用通过双荧光素酶报告基因测定和RIP测定进行验证。
■在OA患者中CircRELL1表达上调,在DMM小鼠和LPS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,并有助于降低OARSI评分。沉默CircRELL1可增加体内和体外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,导致抑制炎症因子的产生,细胞凋亡,和ECM降解,从而抑制OA的进展。
■CircRELL1可能通过调节miR-200c-3p促进OA的进展。
UNASSIGNED: 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.
UNASSIGNED: 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.
UNASSIGNED: 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 in vivo and in vitro 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.
UNASSIGNED: CircRELL1 may promote the progression of OA by regulating the miR-200c-3p.