Joo Seong-Soo, Kang Hee-Cheol, Lee Min-Won, Choi Young-Wook, Lee Do-Ik
Department of Immunology, College of Pharmacy, Chung-Ang University, Seoul 156-756, Korea.
Arch Pharm Res. 2003 Dec;26(12):1029-35. doi: 10.1007/BF02994754.
Osteoporosis is recognized as one of the major hormonal deficiency diseases, especially in menopausal women and the elderly. When estrogen is reduced in the body, local factors such as IL-1beta and IL-6, which are known to be related with bone resorption, are increased and promote osteoclastogenesis, which is responsible for bone resorption. In the present study, we investigated whether glucosidic isoflavones (Isocal, PIII) extracted from Sophorae fructus affect the proliferation of osteoblasts and prevent osteoclastogenesis in vitro by attenuating upstream cytokines such as IL-1beta and IL-6 in a human osteoblastic cell line (MG-63) and in a primary osteoblastic culture from SD rat femurs. Interestingly, IL-1beta and IL-6 mRNA were significantly suppressed in osteoblast-like cells treated with 17beta-estradiol (E2) and PIII when compared to positive control (SDB), and this suppression was more effective at 10(-8)% than at the highest concentration of 10(-4)%. In addition, these were confirmed in protein levels using ELISA assay. In the cell line, the cells showed that E2 was the most effective in osteoblastic proliferation over the whole range of concentration (10(-4)%-10(-12)%), even though PIII also showed the second greatest effectiveness at 10(-8)%. Nitric oxide (NO) was significantly (p<0.05) upregulated in PIII and E2 over the concentration range 10(-6)% to 10(-8)% when compared to SDB, without showing any dose dependency. In bone marrow primary culture, we found by TRAP assay that PIII effectively suppressed osteoclastogenesis next to E2 in comparison with SDB and culture media (control). In conclusion, these results suggest that local bone-resorbing cytokines can be regulated by PIII at lower concentrations and that, therefore, PIII may preferentially induce anti-osteoporosis response by attenuating osteoclastic differentiation and by upregulating NO.
骨质疏松症被认为是主要的激素缺乏疾病之一，尤其是在绝经后女性和老年人中。当体内雌激素减少时，已知与骨吸收相关的局部因子如白细胞介素-1β（IL-1β）和白细胞介素-6（IL-6）会增加，并促进破骨细胞生成，而破骨细胞生成负责骨吸收。在本研究中，我们调查了从槐角中提取的糖苷异黄酮（异卡洛，PIII）是否会影响成骨细胞的增殖，并通过在人成骨细胞系（MG-63）和SD大鼠股骨原代成骨细胞培养物中减弱上游细胞因子如IL-1β和IL-6来体外预防破骨细胞生成。有趣的是，与阳性对照（SDB）相比，用17β-雌二醇（E2）和PIII处理的成骨样细胞中IL-1β和IL-6 mRNA显著受到抑制，并且这种抑制在10^(-8)%时比最高浓度10^(-4)%时更有效。此外，使用酶联免疫吸附测定法在蛋白质水平上证实了这些结果。在细胞系中，细胞显示E2在整个浓度范围（10^(-4)% - 10^(-12)%）内对成骨细胞增殖最有效，尽管PIII在10^(-8)%时也显示出第二大有效性。与SDB相比，在10^(-6)%至10^(-8)%的浓度范围内，PIII和E2中的一氧化氮（NO）显著（p<0.05）上调，且未显示出任何剂量依赖性。在骨髓原代培养中，通过抗酒石酸酸性磷酸酶（TRAP）测定，我们发现与SDB和培养基（对照）相比，PIII在E2之后有效地抑制了破骨细胞生成。总之，这些结果表明局部骨吸收细胞因子可以在较低浓度下被PIII调节，因此，PIII可能通过减弱破骨细胞分化和上调NO来优先诱导抗骨质疏松反应。
