背景:尽管已经报道了环状RNA(circularRNAs,circRNAs)与多种病理状况进展的关系,骨关节炎(OA)中的circRNA参与者几乎没有研究。
方法:在本研究中,我们招募了25例接受关节成形术的OA患者进行软骨组织收集.检索来自基因表达Omnibus的公共circRNA微阵列数据用于circRNA鉴定。通过用IL-1β处理人软骨细胞(CHON-001细胞系),构建了OA相关损伤的体外细胞模型,用circSOD2siRNA沉默circSOD2表达,研究其在细胞凋亡中的作用,炎症反应,和细胞外基质(ECM)降解。此外,我们调查了circSOD2,miR-224-5p,和过氧化物酶3(PRDX3)通过荧光素酶报告基因测定,RNA免疫沉淀测定,和定量逆转录聚合酶链反应。
结果:我们的发现揭示了circSOD2在OA软骨和细胞样品中的过度表达,和circSOD2敲除减轻ECM降解,炎症,CHON-001细胞模型中的凋亡。此外,我们的研究结果表明circSOD2敲低对miR-224-5p表达的调节功能,而miR-224-5p能够下调PRDX3表达。miR-224-5p抑制剂或pcDNA-PRDX3的共转染可以防止circSOD2敲低的作用。
结论:因此,我们的结果表明,circSOD2敲低可作为通过调节miR-224-5p/PRDX3信号轴缓解OA进展的干预策略.
BACKGROUND: Although the implications of circular RNAs (circRNAs) with the progression of diverse pathological conditions have been reported, the circRNA players in osteoarthritis (OA) are barely studied.
METHODS: In this study, twenty-five OA patients who received arthroplasty were recruited for cartilage tissue collection. Public circRNA microarray data from Gene Expression Omnibus was retrieved for circRNA identification. An in vitro cell model of OA-related damages was constructed by treating human chondrocytes (CHON-001 cell line) with IL-1β, and circSOD2 siRNA was used to silence circSOD2 expression to study its functional role in apoptosis, inflammatory responses, and extracellular matrix (ECM) degradation. Besides, we investigated the functional interactions among circSOD2, miR-224-5p, and peroxiredoxin 3 (PRDX3) by luciferase reporter assay, RNA-immunoprecipitation assay, and quantitative reverse transcription polymerase chain reaction.
RESULTS: Our findings revealed the overexpression of circSOD2 in the OA cartilage and cell samples, and circSOD2 knockdown alleviated ECM degradation, inflammation, and apoptosis in CHON-001 cell model. In addition, our findings suggested the regulatory function of circSOD2 knockdown on miR-224-5p expression, while miR-224-5p was capable of downregulating PRDX3 expression. The co-transfection of miR-224-5p inhibitor or pcDNA-PRDX3 could prevent the effect of circSOD2 knockdown.
CONCLUSIONS: Hence, our results demonstrated that knockdown of circSOD2 may serve as an intervention strategy to alleviate OA progression through modulating miR-224-5p/PRDX3 signaling axis.