Department of Orthopedic Surgery, Faculty of Medicine, and Graduate School of Medicine, Hokkaido University, Sapporo, Japan.
Arthritis Rheumatol. 2023 Aug;75(8):1358-1369. doi: 10.1002/art.42505. Epub 2023 Jun 4.
The severity of osteoarthritis (OA) and cartilage degeneration is highly correlated with the development of synovitis, which is mediated by the activity of inflammatory macrophages. A better understanding of intercellular communication between inflammatory macrophages and chondrocytes should aid in the discovery of novel therapeutic targets. We undertook this study to explore the pathologic role of inflammatory macrophage extracellular vesicles (EVs) in cartilage degeneration.
Macrophages were stimulated by treatment with bacterial lipopolysaccharides to mimic the state of inflammatory macrophages, and the resulting EVs were harvested for chondrocyte stimulation in vitro and for intraarticular injection in a mouse model. The stimulated chondrocytes were further subjected to RNA-sequencing analysis and other functional assays. The action of caspase 11 was disrupted in vitro using a specific small interfering RNA or wedelolactone, and in experimental murine OA models by intraarticular injection of wedelolactone.
Stimulated chondrocytes exhibited a significant elevation in the expression of chondrocyte catabolic factors. Consistent with these results, RNA-sequencing analyses of stimulated chondrocytes indicated that up-regulated genes were mainly categorized into apoptotic process and tumor necrosis factor signaling pathways, which suggests the induction of apoptotic process. Moreover, these chondrocytes exhibited a significant elevation in the expression of pyroptosis-related molecules that were correlated with the expression of chondrocyte catabolic factors. The disruption of caspase 11 significantly alleviated pyroptotic and catabolic processes in stimulated chondrocytes and pathologic changes in collagenase-induced and joint instability-induced OA models.
Our results provide new insight into the pathologic mechanisms of OA and suggest that noncanonical pyroptosis in chondrocytes represents an attractive therapeutic target for treatment.
骨关节炎(OA)的严重程度和软骨退化与滑膜炎的发展高度相关,滑膜炎是由炎症巨噬细胞的活性所介导的。更好地了解炎症巨噬细胞和软骨细胞之间的细胞间通讯,应该有助于发现新的治疗靶点。我们进行这项研究是为了探索炎症巨噬细胞细胞外囊泡(EVs)在软骨退化中的病理作用。
用细菌脂多糖刺激巨噬细胞,模拟炎症巨噬细胞的状态,然后提取其 EVs,用于体外刺激软骨细胞,并在小鼠模型中进行关节内注射。进一步对受刺激的软骨细胞进行 RNA 测序分析和其他功能测定。体外使用特异性小干扰 RNA 或 wedelolactone 破坏 caspase 11 的作用,并用 wedelolactone关节内注射破坏实验性小鼠 OA 模型中的 caspase 11 作用。
受刺激的软骨细胞中软骨细胞分解代谢因子的表达显著升高。与这些结果一致,受刺激的软骨细胞的 RNA 测序分析表明,上调的基因主要分为凋亡过程和肿瘤坏死因子信号通路,这表明诱导了凋亡过程。此外,这些软骨细胞中与软骨细胞分解代谢因子表达相关的细胞焦亡相关分子的表达显著升高。caspase 11 的阻断显著缓解了受刺激的软骨细胞中的细胞焦亡和分解代谢过程,以及胶原酶诱导和关节不稳定诱导的 OA 模型中的病理变化。
我们的结果为 OA 的病理机制提供了新的见解,并表明软骨细胞中的非经典细胞焦亡是一种有吸引力的治疗靶点。