Sun Wei, Lv Zhongyang, Li Weitong, Lu Jun, Xie Ya, Wang Peng, Jiang Ruiyang, Dong Jian, Guo Hu, Liu Zizheng, Fei Yuxiang, Tan Guihua, Wang Maochun, Ren Kewei, Xu Jun, Sun Huiqing, Jiang Xuefeng, Shi Dongquan
Department of Orthopedics, Jiangyin People's Hospital Affiliated to Nantong University, 163 Shoushan Road, Jiangyin, 214400, Jiangsu, PR China.
State Key Laboratory of Pharmaceutical Biotechnology, Division of Sports Medicine and Adult Reconstructive Surgery, Department of Orthopedic Surgery, Affiliated Drum Tower Hospital, Medical School, Nanjing University, 321 Zhongshan Road, Nanjing, 210008, Jiangsu, PR China.
J Orthop Translat. 2024 Jan 22;44:114-124. doi: 10.1016/j.jot.2023.12.005. eCollection 2024 Jan.
Osteoarthritis (OA) is the most common age-related musculoskeletal disease. However, there is still a lack of therapy that can modify OA progression due to the complex pathogenic mechanisms. The aim of the study was to explore the role and mechanism of XJB-5-131 inhibiting chondrocytes ferroptosis to alleviate OA progression.
We treated tert-butyl hydroperoxide (TBHP)-induced ferroptosis of mouse primary chondrocytes with XJB-5-131 in vitro. The intracellular ferroptotic hallmarks, cartilage anabolic and catabolic markers, ferroptosis regulatory genes and proteins were detected. Then we established a mouse OA model via destabilization of the medial meniscus (DMM) surgery. The OA mice were treated with intra-articular injection of XJB-5-131 regularly (2 μM, 3 times per week). After 4 and 8 weeks, we performed micro-CT and histological examination to evaluate the protection role of XJB-5-131 in mouse OA subjects. RNA sequencing analysis was performed to unveil the key downstream gene of XJB-5-131 exerting the anti-ferroptotic effect in OA.
XJB-5-131 significantly suppressed TBHP-induced increases of ferroptotic hallmarks (ROS, lipid peroxidation, and Fe accumulation), ferroptotic drivers (Ptgs2, Pgd, Tfrc, Atf3, Cdo1), while restored the expression of ferroptotic suppressors (Gpx4, Fth1). XJB-5-131 evidently promoted the expression of cartilage anabolic and decreased the expression of cartilage catabolic markers. Moreover, intra-articular injection of XJB-5-131 significantly inhibited the expression of Cox2 and Mmp13, while promoted the expression of Col2a1, Gpx4 and Fth1 in DMM-induced mouse articular cartilage. Further, we identified Pebp1 as a potential target of XJB-5-131 by RNA sequencing analysis. The anti-ferroptosis and chondroprotective effects of XJB-5-131 were significantly diminished by Locostatin, a specific antagonist of Pebp1.
XJB-5-131 significantly protects chondrocytes from ferroptosis in TBHP-induced mouse primary chondrocytes and DMM surgery-induced OA mice model via restoring the expression of Pebp1. XJB-5-131 is a potential therapeutic drug in the management of OA progression.
骨关节炎（OA）是最常见的与年龄相关的肌肉骨骼疾病。然而，由于其复杂的致病机制，目前仍缺乏能够改变OA进展的治疗方法。本研究旨在探讨XJB-5-131抑制软骨细胞铁死亡以减轻OA进展的作用及机制。
我们在体外使用XJB-5-131处理叔丁基过氧化氢（TBHP）诱导的小鼠原代软骨细胞铁死亡。检测细胞内铁死亡标志物、软骨合成和分解代谢标志物、铁死亡调节基因和蛋白质。然后通过内侧半月板不稳定（DMM）手术建立小鼠OA模型。对OA小鼠定期进行关节腔内注射XJB-5-131（2 μM，每周3次）。4周和8周后，进行显微CT和组织学检查，以评估XJB-5-131对小鼠OA模型的保护作用。进行RNA测序分析，以揭示XJB-5-131在OA中发挥抗铁死亡作用的关键下游基因。
XJB-5-131显著抑制TBHP诱导的铁死亡标志物（ROS、脂质过氧化和铁积累）、铁死亡驱动因子（Ptgs2、Pgd、Tfrc、Atf3、Cdo1）的增加，同时恢复铁死亡抑制因子（Gpx4、Fth1）的表达。XJB-5-131明显促进软骨合成标志物的表达，并降低软骨分解代谢标志物的表达。此外，关节腔内注射XJB-5-131显著抑制DMM诱导的小鼠关节软骨中Cox2和Mmp13的表达，同时促进Col2a1、Gpx4和Fth1的表达。此外，通过RNA测序分析，我们确定Pebp1是XJB-5-131的潜在靶点。Pebp1的特异性拮抗剂Locostatin显著减弱了XJB-5-131的抗铁死亡和软骨保护作用。
XJB-5-131通过恢复Pebp1的表达，在TBHP诱导的小鼠原代软骨细胞和DMM手术诱导的OA小鼠模型中显著保护软骨细胞免受铁死亡。XJB-5-131是一种潜在的用于控制OA进展的治疗药物。
