Department of Histology and Embryology, Anhui Medical University, No. 81 Meishan Road, Hefei, 230032, Anhui Province, China.
Department of Human Anatomy, Histology and Embryology, Anhui Medical College, No. 632 Furong Road, Hefei, 230601, Anhui Province, China.
Arthritis Res Ther. 2023 Jun 20;25(1):106. doi: 10.1186/s13075-023-03087-7.
Long-stranded non-coding RNA TUG1 is lowly expressed in osteoarthritic chondrocytes. This study aimed to elucidate the role of TUG1 in osteoarthritic cartilage damage and the underlying mechanisms.
Combined database analysis, using primary chondrocytes as well as the C28/I2 cell line, was performed by qRT-PCR, Western blotting, and immunofluorescence to determine the expression of TUG1, miR-144-3p, DUSP1, and other target proteins. Dual luciferase reporter gene and RIP to verify direct interaction of TUG1 with miR-144-3-p and miR-144-3-p with DUSP1, Annexin V-FITC/PI double staining to detect apoptosis. CCK-8 to detect cell proliferation. The biological significance of TUG1, miR-144-3p, and DUSP1 was assessed in vitro experiments using siRNA for TUG1, mimic and repressor for miR-144-3p, and overexpression plasmid for DUSP1. In this study, all data were subjected to a t-test or one-way analysis of variance with a p-value < 0.05 as the cutoff.
TUG1 expression was closely associated with osteoarthritic chondrocyte damage, and knockdown of TUG1 significantly promoted chondrocyte apoptosis and inflammation. In the present study, we found that TUG1 inhibited chondrocyte apoptosis and inflammation by competitively binding miR-144-3p, deregulating the negative regulatory effect of miR-144-3p on DUSP1, promoting DUSP1 expression, and inhibiting the p38 MAPK signaling pathway.
In conclusion, our study clarifies the role of the ceRNA regulatory network of TUG1/miR-144-3p/DUSP1/P38 MAPK in OA cartilage injury and provides an experimental and theoretical basis for genetic engineering tools to promote articular cartilage repair.
长链非编码 RNA TUG1 在骨关节炎软骨细胞中低表达。本研究旨在阐明 TUG1 在骨关节炎软骨损伤中的作用及其潜在机制。
采用 qRT-PCR、Western blot 和免疫荧光法，结合数据库分析，检测 TUG1、miR-144-3p、DUSP1 等靶蛋白在原代软骨细胞和 C28/I2 细胞系中的表达。双荧光素酶报告基因和 RIP 实验验证 TUG1 与 miR-144-3p 及 miR-144-3p 与 DUSP1 的直接相互作用，Annexin V-FITC/PI 双染检测细胞凋亡，CCK-8 检测细胞增殖。采用 TUG1 siRNA、miR-144-3p 模拟物和抑制剂以及 DUSP1 过表达质粒，在体外实验中评估 TUG1、miR-144-3p 和 DUSP1 的生物学意义。本研究中，所有数据均采用 t 检验或单因素方差分析，p 值<0.05 为差异有统计学意义。
TUG1 表达与骨关节炎软骨细胞损伤密切相关，下调 TUG1 可显著促进软骨细胞凋亡和炎症。本研究发现，TUG1 通过竞争性结合 miR-144-3p 抑制软骨细胞凋亡和炎症，解除 miR-144-3p 对 DUSP1 的负调控作用，促进 DUSP1 表达，抑制 p38 MAPK 信号通路。
综上所述，本研究阐明了 TUG1/miR-144-3p/DUSP1/P38 MAPK 这一 ceRNA 调控网络在 OA 软骨损伤中的作用，为促进关节软骨修复的基因工程工具提供了实验和理论依据。
