Hsa_circ_0004662 通过 microRNA-424-5p/VEGFA 轴加速骨关节炎进展。
Hsa_circ_0004662 Accelerates the Progression of Osteoarthritis the microRNA-424-5p/VEGFA Axis.
机构信息
Department of Orthopedics, The First Affiliated Hospital of Shenzhen University/Shenzhen Second People's Hospital, ShenZhen, 518000, P.R. China.
Department of Orthopedics, the First Affiliated Hospital of Shenzhen University/Shenzhen Second People's Hospital, ShenZhen, 518000, P.R. China.
出版信息
Curr Mol Med. 2024;24(2):217-225. doi: 10.2174/1566524023666221103161203.
OBJECTIVE
Circular RNAs (circRNAs) have been extensively implicated in osteoarthritis (OA) progression. Therefore, this study explores the impact of hsa_circ_0004662 on OA progression and the related molecular mechanism.
METHODS
Human articular chondrocyte injury was induced by IL-1β to construct the OA model in vitro. Hsa_circ_0004662 and microRNA (miR)-424-5p expression in chondrocytes was evaluated with qRT-PCR. Vascular endothelial growth factors A (VEGFA) expression was examined with qRT-PCR and western blot after hsa_circ_0004662 knockdown or miR-424-5p overexpression in chondrocytes. Subsequent to loss- and gain-of-function assays in IL-1β-induced chondrocytes, the proliferation and apoptosis of chondrocytes were assessed with CCK-8 assay and flow cytometry, respectively. The expression of MMP13, Aggrecan, and apoptosis-related proteins Bax and Bcl-2 was measured with western blot. The binding of miR-424-5p to hsa_circ_0004662 and VEGFA was assessed with a dual-luciferase reporter gene assay.
RESULTS
Hsa_circ_0004662 was up-regulated, but miR-424-5p was down-regulated in IL-1β-induced chondrocytes. Mechanistically, both hsa_circ_0004662 and VEGFA bound to miR-424-5p, and hsa_circ_0004662 enhanced VEGFA expression by downregulating miR-424-5p. Hsa_circ_0004662 knockdown elevated cell proliferation, decreased apoptosis and MMP13 and Bax expression, and increased Aggrecan and Bcl- 2 expression in IL-1β-induced chondrocytes, which was counteracted by further miR- 424-5p down-regulation or VEGFA overexpression.
CONCLUSION
Hsa_circ_0004662 facilitates OA progression via the miR-424-5p/ VEGFA axis.
目的
环状 RNA（circRNAs）在骨关节炎（OA）进展中广泛涉及。因此，本研究探讨了 hsa_circ_0004662 对 OA 进展的影响及其相关分子机制。
方法
用白细胞介素-1β（IL-1β）诱导人关节软骨细胞损伤，构建体外 OA 模型。用 qRT-PCR 评估软骨细胞中 hsa_circ_0004662 和 microRNA（miR）-424-5p 的表达。用 qRT-PCR 和 Western blot 检测软骨细胞中 hsa_circ_0004662 敲低或 miR-424-5p 过表达后血管内皮生长因子 A（VEGFA）的表达。在 IL-1β诱导的软骨细胞中进行缺失和功能获得实验后，用 CCK-8 法和流式细胞术分别评估软骨细胞的增殖和凋亡。用 Western blot 检测 MMP13、聚集蛋白聚糖和凋亡相关蛋白 Bax 和 Bcl-2 的表达。用双荧光素酶报告基因实验检测 miR-424-5p 与 hsa_circ_0004662 和 VEGFA 的结合。
结果
IL-1β诱导的软骨细胞中 hsa_circ_0004662 上调，miR-424-5p 下调。机制上，hsa_circ_0004662 和 VEGFA 均与 miR-424-5p 结合，hsa_circ_0004662 通过下调 miR-424-5p 增强 VEGFA 表达。hsa_circ_0004662 敲低可增加 IL-1β诱导的软骨细胞的增殖，减少凋亡和 MMP13 和 Bax 的表达，增加 Aggrecan 和 Bcl-2 的表达，进一步下调 miR-424-5p 或过表达 VEGFA 可拮抗上述作用。
结论
hsa_circ_0004662 通过 miR-424-5p/VEGFA 轴促进 OA 进展。
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