Sakai Eiko, Aoki Yuri, Yoshimatsu Masako, Nishishita Kazuhisa, Iwatake Mayumi, Fukuma Yutaka, Okamoto Kuniaki, Tanaka Takashi, Tsukuba Takayuki
Division of Dental Pharmacology, Department of Developmental and Reconstructive Medicine, Nagasaki University Graduate School of Biomedical Sciences, Nagasaki 852-8588, Japan.
Division of Dental Pharmacology, Department of Developmental and Reconstructive Medicine, Nagasaki University Graduate School of Biomedical Sciences, Nagasaki 852-8588, Japan.
Phytomedicine. 2016 Jul 15;23(8):828-37. doi: 10.1016/j.phymed.2016.04.002. Epub 2016 Apr 23.
Osteoclasts are multinucleated bone-resorbing cells that differentiate in response to receptor activator of nuclear factor-κB (NF-κB) ligand (RANKL). Enhanced osteoclastogenesis contributes to bone diseases, such as osteoporosis and rheumatoid arthritis. Rubus parvifolius L. is traditionally used as an herbal medicine for rheumatism; however, its detailed chemical composition and the molecular mechanisms responsible for its biological action have not been elucidated.
To investigate the mechanisms by which R. parvifolius L. extract and its major constituent sanguiin H-6, inhibit osteoclastogenesis and bone resorption.
Cell proliferation, cell differentiation, and bone resorption were detected in vitro. Inhibition of signaling pathways, marker protein expression, and protein nuclear translocation were evaluated by western blot analysis. Tumor necrosis factor-α (TNF-α)-mediated osteoclastogenesis was examined in vivo.
R. parvifolius L. extract inhibited the bone-resorption activity of osteoclasts. In addition, sanguiin H-6 markedly inhibited RANKL-induced osteoclast differentiation and bone resorption, reduced reactive oxygen species production, and inhibited the phosphorylation of inhibitor of NF-κB alpha (IκBα) and p38 mitogen-activated protein kinase. Sanguiin H-6 also decreased the protein levels of nuclear factor of activated T cells cytoplasmic-1 (NFATc1), cathepsin K, and c-Src. Moreover, sanguiin H-6 inhibited the nuclear translocation of NFATc1, c-Fos, and NF-κB in vitro, as well as TNF-α-mediated osteoclastogenesis in vivo.
Our data revealed that R. parvifolius L. has anti-bone resorption activity and suggest that its constituent, sanguiin H-6, can potentially be used for the prevention and treatment of bone diseases associated with excessive osteoclast formation and subsequent bone destruction.
破骨细胞是多核的骨吸收细胞，其分化受核因子κB（NF-κB）受体激活剂配体（RANKL）的影响。破骨细胞生成增强会导致诸如骨质疏松症和类风湿性关节炎等骨疾病。小悬钩子传统上被用作治疗风湿病的草药；然而，其详细的化学成分及其生物学作用的分子机制尚未阐明。
研究小悬钩子提取物及其主要成分山奈酚H-6抑制破骨细胞生成和骨吸收的机制。
在体外检测细胞增殖、细胞分化和骨吸收。通过蛋白质印迹分析评估信号通路的抑制、标志物蛋白表达和蛋白质核转位。在体内检测肿瘤坏死因子-α（TNF-α）介导的破骨细胞生成。
小悬钩子提取物抑制破骨细胞的骨吸收活性。此外，山奈酚H-6显著抑制RANKL诱导的破骨细胞分化和骨吸收，减少活性氧的产生，并抑制NF-κBα抑制剂（IκBα）和p38丝裂原活化蛋白激酶的磷酸化。山奈酚H-6还降低了活化T细胞核因子细胞质1（NFATc1）、组织蛋白酶K和c-Src的蛋白水平。此外，山奈酚H-6在体外抑制NFATc1、c-Fos和NF-κB的核转位，在体内抑制TNF-α介导的破骨细胞生成。
我们的数据表明小悬钩子具有抗骨吸收活性，并表明其成分山奈酚H-6可能可用于预防和治疗与破骨细胞过度形成及随后的骨破坏相关的骨疾病。
