Duan Ao, Ma Zemeng, Shao Xiaolong, Xiong Zhencheng, Zhang Chaoyi, Liu Wenzheng, Wang Guanglin, Hu Shouye, Lin Wei
Department of Orthopedics Surgery, Trauma Medical Centre, West China Hospital, Sichuan University, Chengdu, China.
Department of Orthopedics, Orthopedic Research Institute, West China Hospital, Sichuan University, Chengdu, China.
Clin Transl Med. 2025 Jan;15(1):e70118. doi: 10.1002/ctm2.70118.
Hypoxia can affect the occurrence and development of inflammation in humans, but its effects on the disease progression of osteoarthritis (OA) remain unclear. Synovial macrophages play an essential role in the progression of arthritis. Specifically, the activation of the NOD-like receptor family pyrin domain containing 3 (NLRP3) in macrophages induces the secretion of a series of inflammatory factors, accelerating the progression of OA.
The effects of CBR-470-1 were assessed in a mouse model of OA induced by destabilization of the medial meniscus (DMM) by micro-computed tomography imaging, Safranin-O and Fast Green staining, immunofluorescence staining and enzyme-linked immunosorbent assay. Western Blot analysis was used to explore the underlying mechanism of these experimental results. Additionally, a co-culture system of THP-1 and chondrocytes was established to investigate the impact of CBR-470-1 on chondrocyte proliferation, apoptosis, migration and the regulation of chondrocyte-related proteins within the system.
In hypoxic conditions, CBR-470-1 significantly inhibited the progression of OA in the DMM-induced OA mouse model, but that effect disappeared in the DMM-induced OA phosphoglycerate kinase 1 (PGK1)Lyz2-Cre mouse model. Not only that, CBR-470-1 can also improve the proliferation and migration of chondrocytes, reduce the apoptosis rate of chondrocytes, and regulate the expression of chondrocyte-related proteins in the co-culture system of THP-1 and chondrocytes.
This study conducted a series of in vitro and in vivo experiments, revealing that hypoxia plays a pro-inflammatory role by increasing PGK1 activity and reducing the binding of the deubiquitinating enzyme ubiquitin-specific peptidase 14 to NLRP3, thereby reducing the ubiquitination level of NLRP3. CBR-470-1, a specific inhibitor of PGK1, can reduce PGK1 activity to reverse the role of hypoxia in the progression of OA. These findings lay a foundation for the development of OA treatment in a hypoxic environment.
Hypoxia plays a pro-inflammatory role by increasing PGK1 activity and thereby decreasing the ubiquitination level of NLRP3. Hypoxia plays a pro-inflammatory role by increasing PGK1 activity, reducing the binding of the deubiquitinating enzyme USP14 to NLRP3, and reducing the ubiquitination level of NLRP3. CBR-470-1 reverses the role of hypoxia in the progression of osteoarthritis.
缺氧可影响人类炎症的发生和发展，但其对骨关节炎（OA）疾病进展的影响尚不清楚。滑膜巨噬细胞在关节炎进展中起重要作用。具体而言，巨噬细胞中含吡咯结构域的NOD样受体家族成员3（NLRP3）的激活会诱导一系列炎症因子的分泌，加速OA的进展。
通过微计算机断层扫描成像、番红O和固绿染色、免疫荧光染色和酶联免疫吸附测定，评估CBR - 470 - 1在由内侧半月板不稳定（DMM）诱导的OA小鼠模型中的作用。采用蛋白质免疫印迹分析来探究这些实验结果的潜在机制。此外，建立THP - 1与软骨细胞的共培养系统，以研究CBR - 470 - 1对软骨细胞增殖、凋亡、迁移以及该系统内软骨细胞相关蛋白调控的影响。
在缺氧条件下，CBR - 470 - 1显著抑制DMM诱导的OA小鼠模型中OA的进展，但在DMM诱导的OA磷酸甘油酸激酶1（PGK1）Lyz2 - Cre小鼠模型中该作用消失。不仅如此，CBR - 470 - 1还可改善软骨细胞的增殖和迁移，降低软骨细胞的凋亡率，并在THP - 1与软骨细胞的共培养系统中调节软骨细胞相关蛋白的表达。
本研究进行了一系列体外和体内实验，揭示缺氧通过增加PGK1活性并减少去泛素化酶泛素特异性肽酶14与NLRP3的结合，从而降低NLRP3的泛素化水平，发挥促炎作用。PGK1的特异性抑制剂CBR - 470 - 1可降低PGK1活性，以逆转缺氧在OA进展中的作用。这些发现为缺氧环境下OA治疗的发展奠定了基础。
缺氧通过增加PGK1活性从而降低NLRP3的泛素化水平发挥促炎作用。缺氧通过增加PGK1活性、减少去泛素化酶USP14与NLRP3的结合以及降低NLRP3的泛素化水平发挥促炎作用。CBR - 470 - 1逆转缺氧在骨关节炎进展中的作用。
