Shim Ki-Shuk, Kim Taesoo, Ha Hyunil, Lee Chung-Jo, Lee Bohyoung, Kim Han Sung, Park Ji Hyung, Ma Jin Yeul
Korean Medicine-Based Herbal Drug Development Group, Korea Institute of Oriental Medicine, Daejeon, Korea.
Department of Biomedical Engineering, Institute of Medical Engineering and Yonsei-Fraunhofer Medical Device Lab, Yonsei University, Wonju, Korea.
Integr Med Res. 2015 Jun;4(2):102-111. doi: 10.1016/j.imr.2015.02.002. Epub 2015 Feb 11.
cortex has been traditionally used to treat stomach and intestine diseases in traditional Korean medicine. In this study, we investigated the effect of water extract of cortex (WEMC) on osteoclast differentiation and function.
Phytochemical characterization of WEMC was performed by high-performance liquid chromatography analysis. Osteoclast differentiation of bone marrow-derived macrophages was determined by tartrate-resistant acid phosphatase activity assay. Receptor activator of nuclear factor-κB ligand (RANKL) signaling factors and transcription factors regulating osteoclast differentiation were analyzed by Western blot and real-time polymerase chain reaction. Bone resorption function of mature osteoclasts was examined by using culture plate coated with inorganic crystalline calcium phosphate. Furthermore, the effect of WEMC on osteoporosis was examined using RANKL-induced bone loss model, characterized by micro-computed tomography and bone metabolism marker analysis.
WEMC inhibited RANKL-induced osteoclast differentiation and the bone resorbing activity of mature osteoclasts. WEMC contains gallic acid and honokiol as active constituents contributing to the inhibitory effect of WEMC on osteoclast differentiation. Further, WEMC suppressed RANKL-induced activation of p38 and nuclear factor-κB pathways and expression of osteoclastogenic transcription factors such as c-Fos for AP-1 and nuclear factor of activated T cells cytoplasmic 1. Ectopic overexpression of a constitutive active form of nuclear factor of activated T cells cytoplasmic 1 rescued the antiosteoclastogenic effect of WEMC. Consistent with the results, WEMC suppressed RANKL-induced trabecular bone loss in mice.
WEMC might have a therapeutic potential to treat pathological bone diseases due to increased osteoclast differentiation and function.
在传统韩医学中，皮层传统上用于治疗胃肠疾病。在本研究中，我们研究了皮层水提取物（WEMC）对破骨细胞分化和功能的影响。
通过高效液相色谱分析对WEMC进行植物化学表征。通过抗酒石酸酸性磷酸酶活性测定来确定骨髓来源巨噬细胞的破骨细胞分化。通过蛋白质免疫印迹和实时聚合酶链反应分析调节破骨细胞分化的核因子κB受体激活剂配体（RANKL）信号因子和转录因子。使用涂有无机结晶磷酸钙的培养板检测成熟破骨细胞的骨吸收功能。此外，使用RANKL诱导的骨质流失模型，通过微计算机断层扫描和骨代谢标志物分析，研究WEMC对骨质疏松症的影响。
WEMC抑制RANKL诱导的破骨细胞分化和成熟破骨细胞的骨吸收活性。WEMC含有没食子酸和厚朴酚作为活性成分，有助于WEMC对破骨细胞分化的抑制作用。此外，WEMC抑制RANKL诱导的p38和核因子κB途径的激活以及破骨细胞生成转录因子如AP-1的c-Fos和活化T细胞核因子细胞质1的表达。活化T细胞核因子细胞质1组成型活性形式的异位过表达挽救了WEMC的抗破骨细胞生成作用。与结果一致，WEMC抑制RANKL诱导的小鼠小梁骨丢失。
由于破骨细胞分化和功能增加，WEMC可能具有治疗病理性骨疾病的潜力。
