Jang Ji Yun, Lee Seul Ah, Kim Do Kyung, Lee Sook-Young, Kim Chun Sung
Marine Healthcare Research and Evaluation Center, Chosun University, Wando 59146, Republic of Korea.
Department of Oral Biochemistry, College of Dentistry, Chosun University, Gwangju 61452, Republic of Korea.
Int J Mol Sci. 2024 Dec 13;25(24):13391. doi: 10.3390/ijms252413391.
() was extracted using fermented ethanol. The effect of fermented ethanol extract of (FeCH) on chondrocyte viability was analyzed by 3-(4,5-dimethylthiazol-2-yl)-2,5-iphenyltetrazolium bromide assay, which showed no cytotoxicity at 2 mg/mL. FeCH pretreatment in IL-1β-stimulated chondrocytes significantly inhibited the accumulation of nitric oxide and prostaglandin E, which was analyzed using the ELISA assay. In addition, protein expression levels of inflammatory-related factors, such as inducible nitric oxide synthase, cyclooxygenase-2, interleukin-6, tumor necrosis factor-alpha, and cartilage-degrading-related enzymes, such as matrix metalloproteinases-1, -3, and -13, and a disintegrin and metalloproteinase with thrombospondin motifs-4 and -5 were significantly decreased in IL-1β-stimulated chondrocytes pretreated with FeCH, which were analyzed using western blot analysis. In addition, as a result of analyzing the content of collagen type II (Col II) and proteoglycan through western blot analysis and alcian blue staining, FeCH pretreatment prevented the degradation of Col II and proteoglycan. It was analyzed through western blot analysis that the chondroprotective effect of FeCH may be mediated through MAPKs and NF-κB-signaling mechanisms. In an in vivo study, an osteoarthritis experimental animal model with damaged medial meniscus (DMM) was utilized and orally administered daily for 8 weeks after surgery. At the study end point, knee joints were harvested and subjected to histological analysis with safranin O staining. As a result, articular cartilage was significantly protected in the FeCH group compared to the DMM group. These results suggest FeCH as a candidate material for the development of pharmaceutical materials for the treatment or prevention of degenerative arthritis.
()采用发酵乙醇提取。通过3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四氮唑溴盐法分析了(FeCH)发酵乙醇提取物对软骨细胞活力的影响,结果显示在2mg/mL时无细胞毒性。FeCH对白细胞介素-1β刺激的软骨细胞进行预处理,可显著抑制一氧化氮和前列腺素E的积累,这通过酶联免疫吸附测定法进行分析。此外,在FeCH预处理的白细胞介素-1β刺激的软骨细胞中,炎症相关因子如诱导型一氧化氮合酶、环氧化酶-2、白细胞介素-6、肿瘤坏死因子-α以及软骨降解相关酶如基质金属蛋白酶-1、-3和-13,还有含血小板反应蛋白基序的解聚素和金属蛋白酶-4和-5的蛋白质表达水平显著降低,这通过蛋白质免疫印迹分析进行检测。此外,通过蛋白质免疫印迹分析和阿尔辛蓝染色分析II型胶原蛋白(Col II)和蛋白聚糖的含量,结果显示FeCH预处理可防止Col II和蛋白聚糖的降解。通过蛋白质免疫印迹分析发现,FeCH的软骨保护作用可能通过丝裂原活化蛋白激酶和核因子κB信号传导机制介导。在一项体内研究中,利用内侧半月板损伤(DMM)的骨关节炎实验动物模型,术后每天口服给药8周。在研究终点,采集膝关节并进行番红O染色的组织学分析。结果显示,与DMM组相比,FeCH组的关节软骨得到了显著保护。这些结果表明FeCH可作为开发治疗或预防退行性关节炎药物材料的候选物质。