Abstract：
Previous studies have reported that circular RNA hsa_circ_0010024 (circDHRS3), microRNA (miR)-193a-3p, and Methyl CpG binding protein 2 (MECP2) are unconventionally expressed in osteoarthritis (OA) cartilage samples. However, the regulatory mechanisms among circDHRS3, miR-193a-3p, and MECP2 in OA pathogenesis are unclear. Changes of circDHRS3, miR-193a-3p, and MECP2 mRNA were detected by qRT-PCR. Several protein levels were evaluated using western blotting. Cell proliferation was analyzed by 5-Ethynyl-2\'-deoxyuridine (EdU) and cell counting assays. Cell apoptosis was determined by flow cytometry assay. Detection of pro-inflammatory cytokines was conducted using ELISA. The relationship between circDHRS3 or MECP2 and miR-193a-3p was validated by dual-luciferase reporter assay. We verified that circDHRS3 and MECP2 were overexpressed in OA cartilage samples, whereas miR-193a-3p was downregulated. CircDHRS3 silencing weakened IL-1β-induced chondrocyte cartilage extracellular matrix (ECM) degradation, apoptosis, and inflammatory response. CircDHRS3 adsorbed miR-193a-3p to modulate MECP2 expression. Also, silencing of miR-193a-3p impaired circDHRS3 silencing-mediated suppression on IL-1β-induced chondrocyte injury. Also, MECP2 overexpression alleviated miR-193a-3p mimic-mediated inhibition on IL-1β-prompted chondrocyte injury. CircDHRS3 silencing reduced MECP2 expression via sponging miR-193a-3p, thereby weakening IL-1β-induced chondrocyte ECM degradation, apoptosis, and inflammatory response.
摘要：
以前的研究报道，环状RNAhsa_circ_0010024(circddhrs3)，microRNA(miR)-193a-3p,和甲基CpG结合蛋白2（MECP2）在骨关节炎（OA）软骨样品中非常规表达。然而,circDHRS3、miR-193a-3p、而MECP2在OA发病机制中的作用尚不清楚。circDHRS3,miR-193a-3p,qRT-PCR检测MECP2mRNA。使用蛋白质印迹评价几种蛋白质水平。通过5-乙炔基-2'-脱氧尿苷(EdU)和细胞计数测定分析细胞增殖。通过流式细胞术测定细胞凋亡。使用ELISA进行促炎细胞因子的检测。circirdDHRS3或MECP2与miR-193a-3p之间的关系通过双荧光素酶报告基因测定进行验证。我们验证了circdDHRS3和MECP2在OA软骨样品中过度表达，而miR-193a-3p下调。CircDHRS3沉默减弱IL-1β诱导的软骨细胞软骨细胞外基质（ECM）降解，凋亡，和炎症反应。CircDHRS3吸附miR-193a-3p以调节MECP2表达。此外，miR-193a-3p的沉默损害了circdHRS3沉默介导的对IL-1β诱导的软骨细胞损伤的抑制。此外，MECP2过表达减轻了miR-193a-3p模拟物介导的对IL-1β提示的软骨细胞损伤的抑制。CircDHRS3沉默通过增强miR-193a-3p降低MECP2表达，从而削弱IL-1β诱导的软骨细胞ECM降解，凋亡，和炎症反应。
