目的:本研究的目的是研究ASAVI是否通过调节线粒体功能来控制骨关节炎(OA)。
方法:从大鼠膝关节中分离培养原代软骨细胞。用ASAVI和白细胞介素-1β(IL-1β)处理软骨细胞以模拟OA的炎症环境。细胞活力,凋亡,炎性细胞因子水平,评估细胞外基质(ECM)成分水平。线粒体功能,包括ATP水平,线粒体膜电位,活性氧(ROS)水平,和线粒体DNA含量,进行了评估。Sirtuin3(Sirt3)的表达,线粒体稳态的关键调节因子,被检查过。此外,通过使内侧半月板失稳建立大鼠OA模型,并评估了ASAVI对软骨退变的影响。
结果:ASAVI处理改善了细胞活力,减少细胞凋亡,并降低IL-1β诱导的软骨细胞中IL-6和TNF-α的水平。ASAVI也上调II型胶原和Aggrecan表达,同时下调ADAMTS5和MMP-13的表达。此外,ASAVI通过增加ATP水平减轻IL-1β诱导的线粒体功能障碍,恢复线粒体膜电位,减少ROS的产生,并保留线粒体DNA含量。这些作用伴随着Sirt3的激活。在大鼠OA模型中,ASAVI治疗增加Sirt3表达并减轻软骨退变。
结论:ASAVI通过激活Sirt3改善线粒体功能,对IL-1β诱导的软骨细胞具有软骨保护和抗炎作用。ASAVI还减弱大鼠OA模型中的软骨变性。这些发现表明,ASAVI可能是通过靶向线粒体功能障碍治疗骨关节炎的潜在治疗剂。
OBJECTIVE: The aim of this study was to investigate whether ASA VI controls osteoarthritis (OA) by regulating mitochondrial function.
METHODS: Primary chondrocytes were isolated and cultured from rat knee joints. The chondrocytes were treated with ASA VI and interleukin-1β (IL-1β) to simulate the inflammatory environment of OA. Cell viability, apoptosis, inflammatory cytokine levels, and extracellular matrix (ECM) component levels were assessed. Mitochondrial function, including ATP levels, mitochondrial membrane potential, reactive oxygen species (ROS) levels, and mitochondrial DNA content, was evaluated. The expression of Sirtuin 3 (Sirt3), a key regulator of mitochondrial homeostasis, was examined. Additionally, a rat OA model was established by destabilizing the medial meniscus, and the effects of ASA VI on cartilage degeneration were assessed.
RESULTS: ASA VI treatment improved cell viability, reduced apoptosis, and decreased IL-6 and TNF-α levels in IL-1β-induced chondrocytes. ASA VI also upregulated Collagen II and Aggrecan expression, while downregulating ADAMTS5 and MMP-13 expression. Furthermore, ASA VI mitigated IL-1β-induced mitochondrial dysfunction by increasing ATP levels, restoring mitochondrial membrane potential, reducing ROS production, and preserving mitochondrial DNA content. These effects were accompanied by the activation of Sirt3. In the rat OA model, ASA VI treatment increased Sirt3 expression and alleviated cartilage degeneration.
CONCLUSIONS: ASA VI exerts chondroprotective and anti-inflammatory effects on IL-1β-induced chondrocytes by improving mitochondrial function through Sirt3 activation. ASA VI also attenuates cartilage degeneration in a rat OA model. These findings suggest that ASA VI may be a potential therapeutic agent for the treatment of osteoarthritis by targeting mitochondrial dysfunction.