Department of Orthopedics, The Second Affiliated Hospital of Nanchang University, Donghu District, Nanchang City, Jiangxi Province, China.
Department of Sports Medicine, The Affiliated Ganzhou Hospital of Nanchang University (Ganzhou People's Hospital), Zhanggong District, Ganzhou City, Jiangxi Province, China.
PLoS One. 2024 Apr 9;19(4):e0298575. doi: 10.1371/journal.pone.0298575. eCollection 2024.
Osteoarthritis (OA) is a widespread chronic, progressive, degenerative joint disease that causes pain and disability. Current treatments for OA have limited effectiveness and new biomarkers need to be identified. Bioinformatics analysis was conducted to explore differentially expressed genes and DNA repair/recombination protein 54 L (RAD54L) was selected. We firstly overexpressed RAD54L in interleukin-1β (IL-1β)-induced human articular chondrocytes or in OA rats to investigate its effect on OA. Chondrocyte viability and apoptotic rate were measured by Cell Counting Kit-8 and flow cytometry, respectively. Then we evaluated OA severity in vivo by Hematoxylin-eosin staining and Osteoarthritis Research Society International standards. The expression of inflammatory mediators was tested by enzyme-linked immunosorbent assay. Finally, western blot was performed to determine the relative expression level of hypoxia-inducible factors 1α (HIF-1α) and vascular endothelial growth factor (VEGF). Overexpression of RAD54L promoted cell viability and attenuated apoptosis in IL-1β-induced human chondrocytes. A lower Osteoarthritis Research Society International score and a remarkable alleviation of chondrocyte disordering and infiltration of inflammatory cells were found in cartilage tissues of OA rats after overexpressing RAD54L. The inflammatory response induced by OA was decreased by RAD54L overexpression in vitro and in vivo. In addition, RAD54L overexpression decreased the relative expression level of HIF-1α and VEGF. Overexpression of RAD54L could attenuate OA by suppressing the HIF-1α/VEGF signaling pathway, indicating that RAD54L may be a potential treatment target for OA.
骨关节炎(OA)是一种广泛存在的慢性、进行性、退行性关节疾病,可导致疼痛和残疾。目前 OA 的治疗方法效果有限,需要确定新的生物标志物。本研究采用生物信息学分析方法,筛选差异表达基因,发现 DNA 修复/重组蛋白 54 L(RAD54L)。首先在白细胞介素-1β(IL-1β)诱导的人关节软骨细胞或 OA 大鼠中过表达 RAD54L,探讨其对 OA 的影响。采用细胞计数试剂盒-8 和流式细胞术分别检测软骨细胞活力和凋亡率。然后通过苏木精-伊红染色和骨关节炎研究协会国际标准评估体内 OA 严重程度。采用酶联免疫吸附试验检测炎症介质的表达。最后,采用 Western blot 检测低氧诱导因子 1α(HIF-1α)和血管内皮生长因子(VEGF)的相对表达水平。过表达 RAD54L 可促进 IL-1β诱导的人软骨细胞活力,抑制细胞凋亡。过表达 RAD54L 可降低 OA 大鼠软骨组织的 Osteoarthritis Research Society International 评分,显著减轻软骨细胞紊乱和炎症细胞浸润。体外和体内实验均表明,RAD54L 过表达可减轻 OA 引起的炎症反应。此外,RAD54L 过表达可降低 HIF-1α和 VEGF 的相对表达水平。过表达 RAD54L 可通过抑制 HIF-1α/VEGF 信号通路减轻 OA,表明 RAD54L 可能是 OA 的潜在治疗靶点。