Xu Kanna, Wang Haoran, Wu Zhongqing
Emergency Department, The First People's Hospital of Huzhou, Huzhou, Zhejiang, China.
Department of Orthopedics, Hangzhou Children's Hospital, Hangzhou, Zhejiang, China.
Chin J Physiol. 2023 Jul-Aug;66(4):284-293. doi: 10.4103/cjop.CJOP-D-23-00031.
Osteoarthritis (OA) is a form of chronic degenerative disease contributing to elevated disability rate among the elderly. Genkwanin is an active component extracted from Daphne genkwa possessing pharmacologic effects. Here, this study is designed to expound the specific role of genkwanin in OA and elaborate the probable downstream mechanism. First, the viability of chondrocytes in the presence or absence of interleukin-1 beta (IL-1β) treatment was detected by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay. Terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling assay was used to assess cell apoptosis. Inflammatory response was estimated through enzyme-linked immunosorbent assay and Western blot. In addition, immunofluorescence staining and Western blot were utilized to measure the expression of extracellular matrix (ECM)-associated proteins. Dual-specificity protein phosphatase-1 (DUSP1) expression was tested by reverse transcription-quantitative polymerase chain reaction (RT-qPCR) and Western blot. Following DUSP1 elevation in genkwanin-treated chondrocytes exposed to IL-1β, inflammatory response and ECM-associated factors were evaluated as forementioned. In addition, 5,5',6,6'-tetrachloro-1,1',3,3'-tetraethylbenzimidazolocarbocyanine iodide staining was to assess the mitochondrial membrane potential. Adenosine triphosphate (ATP) level was examined with ATP assay kit, and RT-qPCR was used to test mitochondrial DNA expression. Results indicated that genkwanin administration enhanced the viability while ameliorated the apoptosis, inflammatory response, and ECM degradation in IL-1β-induced chondrocytes. Besides, genkwanin treatment fortified DUSP1 expression in IL-1β-exposed chondrocytes. DUSP1 interference further offsets the impacts of genkwanin on the inflammation, ECM degradation, and mitochondrial dysfunction in IL-1β-challenged chondrocytes. In short, genkwanin enhanced DUSP1 expression to mitigate mitochondrial dysfunction, thus ameliorating IL-1β-elicited inflammation, apoptosis, and degradation of ECM in chondrocytes.
骨关节炎（OA）是一种慢性退行性疾病，导致老年人残疾率升高。芫花素是从芫花中提取的具有药理作用的活性成分。在此，本研究旨在阐述芫花素在骨关节炎中的具体作用，并详细说明可能的下游机制。首先，通过3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四氮唑溴盐法检测白细胞介素-1β（IL-1β）处理或未处理情况下软骨细胞的活力。采用末端脱氧核苷酸转移酶介导的dUTP缺口末端标记法评估细胞凋亡。通过酶联免疫吸附测定和蛋白质印迹法评估炎症反应。此外，利用免疫荧光染色和蛋白质印迹法检测细胞外基质（ECM）相关蛋白的表达。通过逆转录-定量聚合酶链反应（RT-qPCR）和蛋白质印迹法检测双特异性蛋白磷酸酶-1（DUSP1）的表达。在暴露于IL-1β的经芫花素处理的软骨细胞中提高DUSP1表达后，如前所述评估炎症反应和ECM相关因子。此外，用5,5',6,6'-四氯-1,1',3,3'-四乙基苯并咪唑羰花青碘化物染色评估线粒体膜电位。用ATP检测试剂盒检测三磷酸腺苷（ATP）水平，并用RT-qPCR检测线粒体DNA表达。结果表明，给予芫花素可提高IL-1β诱导的软骨细胞的活力，同时改善其凋亡、炎症反应和ECM降解。此外，芫花素处理增强了暴露于IL-1β的软骨细胞中DUSP1的表达。DUSP1干扰进一步抵消了芫花素对IL-1β刺激的软骨细胞炎症、ECM降解和线粒体功能障碍的影响。简而言之，芫花素通过提高DUSP1表达减轻线粒体功能障碍，从而改善IL-1β引起的软骨细胞炎症、凋亡和ECM降解。
