环状 RNA_0003800 通过 miR-197-3p/ SOX5 轴加剧 IL-1β诱导的软骨细胞损伤。

Circular RNA_0003800 exacerbates IL-1β-induced chondrocyte injury via miR-197-3p/SOX5 axis.

机构信息

Department of Orthopedics, The First Affiliated Hospital of Zhengzhou University, China.

Department of Orthopedics, The Affiliated Hospital of Henan University of Chinese Medicine, China.

出版信息

Int Immunopharmacol. 2023 Feb;115:109643. doi: 10.1016/j.intimp.2022.109643. Epub 2023 Jan 5.

DOI:10.1016/j.intimp.2022.109643

PMID:36610331

Abstract

BACKGROUND

Osteoarthritis (OA) is a serious degenerative disease of articular cartilage, which has a great impact on the quality of life of patients. Circular RNA (circRNA) plays an important role in OA progression. Our study aims to explore the role and mechanism of circ_0003800 in OA.

METHODS

Circ_0003800, microRNA-197-3p (miR-197-3p) and SRY-box transcription factor 5 (SOX5) contents were measured by quantitative real-time polymerase chain reaction (qRT-PCR) and western blot. Cell Counting Kit-8 (CCK8), 5-ethynyl-2'-deoxyuridine (EdU), flow cytometry, western blot and enzyme-linked immunosorbent assay (ELISA) were deployed to evaluate cell proliferation, apoptosis, extracellular matrix (ECM) degradation, inflammatory response and oxidative stress. Interaction of miR-197-3p and circ_0003800 or SOX5 was evidenced by dual-luciferase reporter system, RNA immunoprecipitation (RIP) and RNA pull down assays.

RESULTS

OA tissues and model cells had higher abundance of circ_0003800 and SOX5, while miR-197-3p content was lower. Functionally, circ_0003800 knockdown alleviated IL-1β-mediated injury in C28/I2 cells. Mechanistically, circ_0003800 could sponge miR-197-3p, and miR-197-3p could target SOX5. Besides, in-miR-197-3p reversed the suppressive effect of circ_0003800 downregulation on IL-1β-induced C28/I2 cell injury, and SOX5 overexpression could also diminish the inhibitory effect of miR-197-3p on IL-1β-induced C28/I2 cell injury.

CONCLUSION

Circ_0003800 exacerbates IL-1β-induced chondrocyte injury via miR-197-3p/SOX5 axis.

摘要

背景

骨关节炎（OA）是一种严重的关节软骨退行性疾病，对患者的生活质量有很大影响。环状 RNA（circRNA）在 OA 进展中发挥重要作用。我们的研究旨在探讨 circ_0003800 在 OA 中的作用和机制。

方法

通过实时定量聚合酶链反应（qRT-PCR）和 Western blot 测量 circ_0003800、微小 RNA-197-3p（miR-197-3p）和性别决定区 Y 框转录因子 5（SOX5）的含量。细胞计数试剂盒-8（CCK8）、5-乙炔基-2'-脱氧尿苷（EdU）、流式细胞术、Western blot 和酶联免疫吸附测定（ELISA）用于评估细胞增殖、凋亡、细胞外基质（ECM）降解、炎症反应和氧化应激。通过双荧光素酶报告系统、RNA 免疫沉淀（RIP）和 RNA 下拉实验证实 miR-197-3p 与 circ_0003800 或 SOX5 的相互作用。

结果

OA 组织和模型细胞中 circ_0003800 和 SOX5 的含量较高，而 miR-197-3p 的含量较低。功能上，circ_0003800 敲低减轻了 IL-1β 介导的 C28/I2 细胞损伤。机制上，circ_0003800 可以海绵 miR-197-3p，miR-197-3p 可以靶向 SOX5。此外，在 miR-197-3p 逆转了 circ_0003800 下调对 IL-1β 诱导的 C28/I2 细胞损伤的抑制作用，而 SOX5 的过表达也可以减弱 miR-197-3p 对 IL-1β 诱导的 C28/I2 细胞损伤的抑制作用。