He Min, Jia Zhihe, Wen Yiying, Chen Xiaolin
Department of Joint Surgery, Pingxiang People's Hospital, Pingxiang City, 337055, Jiangxi, China.
Department of Laboratory, Pingxiang People's Hospital, No. 8 Wugong Shanzhong Avenue, Development Zone, Pingxiang City, Jiangxi, China.
J Orthop Surg Res. 2022 Mar 24;17(1):177. doi: 10.1186/s13018-022-02970-4.
Osteoarthritis (OA) is a chronic joint disease featured by articular cartilage degeneration and damage. Accumulating evidence have demonstrated the pivotal regulatory roles of circular RNAs in OA pathology. However, the role of circ_0043947 in OA progression and its associated mechanism remain largely unknown.
The expression of RNA and protein was determined by reverse transcription-quantitative polymerase chain reaction and Western blot assay. Cell viability was assessed by 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. Cell proliferation was analyzed by 5-Ethynyl-2'-deoxyuridine (EdU) assay and flow cytometry. Cell apoptosis was assessed by flow cytometry. Enzyme linked immunosorbent assay was conducted to analyze the release of pro-inflammatory cytokines. Dual-luciferase reporter assay and RNA immunoprecipitation assay were performed to confirm the target interaction between microRNA-671-5p (miR-671-5p) and circ_0043947 or reticulon 3 (RTN3).
Interleukin 1β (IL-1β) stimulation up-regulated the expression of circ_0043947 in chondrocytes. IL-1β treatment restrained the viability and proliferation and induced the apoptosis, extracellular matrix degradation and inflammatory response of chondrocytes partly by up-regulating circ_0043947. Circ_0043947 interacted with miR-671-5p, and miR-671-5p silencing largely reversed circ_0043947 knockdown-mediated protective effects in IL-1β-induced chondrocytes. miR-671-5p interacted with the 3' untranslated region (3'UTR) of RTN3. miR-671-5p overexpression attenuated IL-1β-induced injury in chondrocytes, and these protective effects were largely overturned by the overexpression of RTN3. Circ_0043947 acted as a molecular sponge for miR-671-5p to up-regulate RTN3 level in chondrocytes.
Circ_0043947 silencing alleviated IL-1β-induced injury in chondrocytes by targeting miR-671-5p/RTN3 axis.
骨关节炎（OA）是一种以关节软骨退变和损伤为特征的慢性关节疾病。越来越多的证据表明环状RNA在OA病理过程中起关键调节作用。然而，circ_0043947在OA进展中的作用及其相关机制仍不清楚。
通过逆转录定量聚合酶链反应和蛋白质免疫印迹法检测RNA和蛋白质的表达。采用3-(4,5-二甲基噻唑-2)-2,5-二苯基四氮唑溴盐（MTT）法评估细胞活力。通过5-乙炔基-2'-脱氧尿苷（EdU）法和流式细胞术分析细胞增殖。采用流式细胞术评估细胞凋亡。进行酶联免疫吸附测定以分析促炎细胞因子的释放。进行双荧光素酶报告基因测定和RNA免疫沉淀测定以确认微小RNA-671-5p（miR-671-5p）与circ_0043947或网织蛋白3（RTN3）之间的靶向相互作用。
白细胞介素1β（IL-1β）刺激上调软骨细胞中circ_0043947的表达。IL-1β处理部分通过上调circ_0043947抑制软骨细胞的活力和增殖，并诱导软骨细胞凋亡、细胞外基质降解和炎症反应。Circ_0043947与miR-671-5p相互作用，miR-671-5p沉默在很大程度上逆转了circ_0043947敲低介导的对IL-1β诱导的软骨细胞的保护作用。miR-671-5p与RTN3的3'非翻译区（3'UTR）相互作用。miR-671-5p过表达减轻了IL-1β诱导的软骨细胞损伤，而RTN3过表达在很大程度上推翻了这些保护作用。Circ_0043947作为miR-671-5p的分子海绵，上调软骨细胞中RTN3水平。
Circ_0043947沉默通过靶向miR-671-5p/RTN3轴减轻IL-1β诱导的软骨细胞损伤。
