PDF(Pubmed)
Abstract：
This study aimed to investigate whether the beneficial effects of PCA on chondrocyte senescence are mediated through the regulation of mitophagy. Chondrocyte senescence plays a significant role in the development and progression of knee osteoarthritis (OA). The compound protocatechuic aldehyde (PCA), which is abundant in the roots of Salvia miltiorrhiza, has been reported to have antioxidant properties and the ability to protect against cellular senescence. To achieve this goal, a destabilization of the medial meniscus (DMM)-induced mouse OA model and a lipopolysaccharide (LPS)-induced chondrocyte senescence model were used, in combination with PINK1 gene knockdown or overexpression. After treatment with PCA, cellular senescence was assessed using Senescence-Associated β-Galactosidase (SA-β-Gal) staining, DNA damage was evaluated using Hosphorylation of the Ser-139 (γH2AX) staining, reactive oxygen species (ROS) levels were measured using Dichlorodihydrofluorescein diacetate (DCFH-DA) staining, mitochondrial membrane potential was determined using a 5,5\',6,6\'-TETRACHLORO-1,1\',3,3\'-*. TETRAETHYBENZIMIDA (JC-1) kit, and mitochondrial autophagy was examined using Mitophagy staining. Western blot analysis was also performed to detect changes in senescence-related proteins, PINK1/Parkin pathway proteins, and mitophagy-related proteins. Our results demonstrated that PCA effectively reduced chondrocyte senescence, increased the mitochondrial membrane potential, facilitated mitochondrial autophagy, and upregulated the PINK1/Parkin pathway. Furthermore, silencing PINK1 weakened the protective effects of PCA, whereas PINK1 overexpression enhanced the effects of PCA on LPS-induced chondrocytes. PCA attenuates chondrocyte senescence by regulating PINK1/Parkin-mediated mitochondrial autophagy, ultimately reducing cartilage degeneration.
摘要：
本研究旨在探讨PCA对软骨细胞衰老的有益作用是否通过调节线粒体自噬来介导。软骨细胞衰老在膝骨关节炎(OA)的发生、发展中起着重要作用。复方原儿茶醛（PCA），在丹参的根中含量丰富，据报道具有抗氧化性能和防止细胞衰老的能力。为了实现这一目标,使用内侧半月板（DMM）诱导的小鼠OA模型和脂多糖（LPS）诱导的软骨细胞衰老模型，与PINK1基因敲低或过表达相结合。用PCA治疗后，使用衰老相关β-半乳糖苷酶（SA-β-Gal）染色评估细胞衰老，使用Ser-139（γH2AX）染色的Hosphorylation评估DNA损伤，使用二氯二氢荧光素二乙酸酯（DCFH-DA）染色测量活性氧（ROS）水平，线粒体膜电位使用5.5'，6,6'-四氯-1,1',3，3'-*。四苯并咪唑（JC-1）试剂盒，使用线粒体自噬染色检查线粒体自噬。还进行了蛋白质印迹分析以检测衰老相关蛋白的变化，PINK1/Parkin通路蛋白,和线粒体自噬相关蛋白。我们的结果表明，PCA有效地减少软骨细胞衰老，增加了线粒体膜电位，促进线粒体自噬，并上调PINK1/Parkin通路。此外，沉默PINK1削弱了PCA的保护作用，而PINK1过表达增强了PCA对LPS诱导的软骨细胞的作用。PCA通过调节PINK1/Parkin介导的线粒体自噬减弱软骨细胞衰老,最终减少软骨退化。
