Life & Science Research Center, Hyunsung Vital Co. Ltd., Seoul 07255, Korea.
Department of Food and Nutrition, Hannam University, Daejeon 34054, Korea.
Molecules. 2018 Oct 29;23(11):2807. doi: 10.3390/molecules23112807.
Osteoarthritis (OA) is the common form of arthritis and is characterized by disability and cartilage degradation. Although natural product extracts have been reported to have anti-osteoarthritic effects, the potential bioactivity of Ryupunghwan (RPH), a traditional Korean medicinal botanical formula that contains , , , , , , and , is not known well. Therefore, the inhibitory effects of single compounds isolated from RPH on the OA-related molecules were investigated using IL-1β-stimulated chondrosarcoma SW1353 (SW1353) cell model. Two bioactive compounds, isomucronulatol 7--β-d-glucoside (IMG) and ecliptasaponin A (ES) were isolated and purified from RPH using column chromatography, and then the structures were analyzed using ESI-MS, ¹H-NMR, and C-NMR spectrum. The expression or amount of matrix metalloproteinase 13 (MMP13), COX1/2, TNF-α, IL-1β or p65 was determined by RT-PCR, Western blot, and enzyme-linked immunosorbent assay (ELISA). RPH pretreatment reduced the expression and amounts of MMP13, and the expression of collagen II, COX1/2, TNF-α, IL-1β or p65, which were increased in IL-1β-stimulated SW1353 cells. IMG reduced the expression of all OA-related molecules, but the observed inhibitory effect was less than that of RPH extract. The other single compound ES showed the reduced expression of all OA-related molecules, and the effect was stronger than that in IMG (approximately 100 fold). Combination pretreatment of both single components remarkably reduced the expression of MMP13, compared to each single component. These synergic effects may provide potential molecular modes of action for the anti-osteoarthritic effects of RPH observed in clinical and animal studies.
骨关节炎（OA）是关节炎的常见形式，其特征为残疾和软骨降解。虽然已经报道天然产物提取物具有抗骨关节炎作用，但是包含 、 、 、 、 、 和 的传统韩国药用植物配方 Ryupunghwan（RPH）的潜在生物活性尚未得到很好的研究。因此，使用白细胞介素 1β 刺激的软骨肉瘤 SW1353（SW1353）细胞模型研究了从 RPH 中分离的单个化合物对与 OA 相关的分子的抑制作用。使用柱层析从 RPH 中分离和纯化了两种生物活性化合物，异木栓醇 7--β-D-葡萄糖苷（IMG）和旱莲草皂苷 A（ES），然后使用 ESI-MS、1H-NMR 和 C-NMR 谱对其结构进行了分析。通过 RT-PCR、Western blot 和酶联免疫吸附试验（ELISA）测定基质金属蛋白酶 13（MMP13）、COX1/2、TNF-α、IL-1β 或 p65 的表达或量。RPH 预处理可降低 MMP13 的表达和量，以及在白细胞介素 1β 刺激的 SW1353 细胞中增加的胶原 II、COX1/2、TNF-α、IL-1β 或 p65 的表达。IMG 降低了所有与 OA 相关的分子的表达，但观察到的抑制作用小于 RPH 提取物。另一种单一化合物 ES 显示所有与 OA 相关的分子的表达减少，其作用强于 IMG（约 100 倍）。与单一成分相比，两种单一成分的组合预处理可显著降低 MMP13 的表达。这些协同作用可能为临床和动物研究中观察到的 RPH 的抗骨关节炎作用提供潜在的分子作用模式。
