Busa Prabhakar, Lee Sing-Ong, Huang Niancih, Kuthati Yaswanth, Wong Chih-Shung
Department of Anesthesiology, Cathay General Hospital, Taipei City 106, Taiwan.
Department of Anesthesiology, Tri-Service General Hospital, Taipei City 114, Taiwan.
Antioxidants (Basel). 2022 Jun 20;11(6):1209. doi: 10.3390/antiox11061209.
The most common joint disease in the elderly is knee osteoarthritis (OA). It is distinguished by cartilage degradation, subchondral bone loss, and a decrease in joint space. We studied the effects of carnosine (CA) on knee OA in male Wistar rats. OA is induced by anterior cruciate ligament transection combined with medial meniscectomy (ACLT+MMx) method and in vitro studies are conducted in fibroblast-like synoviocyte cells (FLS). The pain was assessed using weight-bearing and paw-withdrawal tests. CA supplementation significantly reduced pain. The enzyme-linked immunosorbent assay (ELISA) method was used to detect inflammatory proteins in the blood and intra-articular synovial fluid (IASF), and CA reduced the levels of inflammatory proteins. Histopathological studies were performed on knee-tissue samples using toluidine blue and hematoxylin and eosin (H and E) assays. CA treatment improved synovial protection and decreased cartilage degradation while decreasing zonal depth lesions. Furthermore, Western blotting studies revealed that the CA-treated group activated nuclear factor erythroid 2-related factor (Nrf2) and heme oxygenase (HO-1) and reduced the expression of cyclooxygenase-2 (COX-2). FLS cells were isolated from the knee joints and treated with IL-1β to stimulate the inflammatory response and increase reactive oxygen species (ROS). The matrix metalloproteinase protein (MMP's) levels (MMP-3, and MMP-13) were determined using the reverse transcription-polymerase chain reaction (RT-PCR), and CA treatment reduced the MMP's expression levels. When tested using the 2',7'-dicholorodihydrofluroscene diacetate (DCFDA) assay and the 5,5',6,6'-tetracholoro-1,1',3,3'-tertraethylbenzimidazolcarboc janine iodide (JC-1) assay in augmented ROS FLS cells, CA reduced the ROS levels and improved the mitochondrial membrane permeability. This study's investigation suggests that CA significantly alleviates knee OA both in vitro and in vivo.
老年人中最常见的关节疾病是膝骨关节炎（OA）。其特征为软骨降解、软骨下骨丢失以及关节间隙变窄。我们研究了肌肽（CA）对雄性Wistar大鼠膝骨关节炎的影响。采用前交叉韧带横断联合内侧半月板切除术（ACLT+MMx）方法诱导骨关节炎，并在成纤维样滑膜细胞（FLS）中进行体外研究。使用负重和爪撤离试验评估疼痛。补充CA可显著减轻疼痛。采用酶联免疫吸附测定（ELISA）法检测血液和关节内滑液（IASF）中的炎症蛋白，CA可降低炎症蛋白水平。使用甲苯胺蓝以及苏木精和伊红（H&E）染色法对膝关节组织样本进行组织病理学研究。CA治疗改善了滑膜保护，减少了软骨降解，同时减少了带状深度病变。此外，蛋白质印迹研究表明，CA治疗组激活了核因子红细胞2相关因子（Nrf2）和血红素加氧酶（HO-1），并降低了环氧化酶-2（COX-2）的表达。从膝关节分离出FLS细胞，用白细胞介素-1β处理以刺激炎症反应并增加活性氧（ROS）。使用逆转录-聚合酶链反应（RT-PCR）测定基质金属蛋白酶蛋白（MMP）水平（MMP-3和MMP-13），CA治疗降低了MMP的表达水平。在增强ROS的FLS细胞中使用2',7'-二氯二氢荧光素二乙酸酯（DCFDA）测定法和5,5',6,6'-四氯-1,1',3,3'-四乙基苯并咪唑羰花青碘化物（JC-1）测定法进行测试时，CA降低了ROS水平并改善了线粒体膜通透性。本研究调查表明，CA在体外和体内均能显著减轻膝骨关节炎。
