Oriental Medicine Research Center for Bone & Joint Disease, KyungHee University, 149, Sangil-dong, Kangdong-gu, Seoul, 134-727, Republic of Korea.
Int Immunopharmacol. 2010 Apr;10(4):500-7. doi: 10.1016/j.intimp.2010.01.012. Epub 2010 Feb 4.
Osteoarthritis is a multifactorial disease characterized by loss of articular cartilage and subchondral plate thickening. Therefore, biochemical analysis of the underlying bone tissue has provided important information for treatment of osteoarthritis. In this study, we determined the potential role of formononetin, a phytoestrogen isolated from Astragalus membranaceus to alter the expression of metabolic markers and cytokine production of human normal osteoblasts (Obs) and osteoarthritis subchondral osteoblasts (OA Obs). Human OA Obs and normal Obs were cultured for 3days, 7days or 14days in the present medium only or were treated with various doses of formononetin. Cells were analyzed for viability by WST-8 assay, alkaline phosphatase (ALP) activity, osteogenic markers (osteocalcin (OCN) and type I collagen (Col I)) and cytokines (interleukin-6 (IL-6), vascular endothelial growth factor (VEGF), bone morphogenic protein-2 (BMP-2)). The level of IL-6, VEGF, BMP-2, OCN and Col I was increased in OA Obs compared with normal Obs. Formononetin dose-dependently decreased ALP, IL-6, VEGF, BMP-2, OCN and Col I in OA Obs, while markedly increased ALP, VEGF, BMP-2, OCN and Col I in normal Obs. Interestingly, formononetin markedly increased the expression of VEGF and BMP-2 for 3days of culture and significantly increased OCN and Col I at 14days in human normal Obs. The remodeling effect of formononetin on osteogenic markers and cytokines of inflammatory mediators was more striking in OA Obs as well. Taken together, these results could suggest that formononetin has biphasic positive effects on normal Obs and OA Obs by modifying their biological synthetic capacities.
骨关节炎是一种多因素疾病，其特征为关节软骨丧失和软骨下板增厚。因此，对潜在骨组织的生化分析为骨关节炎的治疗提供了重要信息。在这项研究中，我们确定了芒柄花素（一种从黄芪中分离出的植物雌激素）改变人正常成骨细胞（Obs）和骨关节炎软骨下成骨细胞（OA Obs）代谢标志物表达和细胞因子产生的潜在作用。将人 OA Obs 和正常 Obs 在本培养基中培养 3 天、7 天或 14 天，或用不同剂量的芒柄花素处理。通过 WST-8 测定法、碱性磷酸酶（ALP）活性、成骨标志物（骨钙素（OCN）和 I 型胶原（Col I））和细胞因子（白细胞介素-6（IL-6）、血管内皮生长因子（VEGF）、骨形态发生蛋白-2（BMP-2））分析细胞活力。与正常 Obs 相比，OA Obs 中的 IL-6、VEGF、BMP-2、OCN 和 Col I 水平升高。芒柄花素呈剂量依赖性地降低 OA Obs 中的 ALP、IL-6、VEGF、BMP-2、OCN 和 Col I，而明显增加正常 Obs 中的 ALP、VEGF、BMP-2、OCN 和 Col I。有趣的是，芒柄花素在培养的第 3 天显著增加了 VEGF 和 BMP-2 的表达，并在第 14 天显著增加了人正常 Obs 中的 OCN 和 Col I。芒柄花素对成骨标志物和炎症介质细胞因子的重塑作用在 OA Obs 中更为明显。总之，这些结果表明，芒柄花素通过改变正常 Obs 和 OA Obs 的生物合成能力，对它们具有双相的积极作用。
