Kim Jeong-Mi, Lee Jung-Han, Lee Guem-San, Noh Eun-Mi, Song Hyun-Kyung, Gu Dong Ryun, Kim Seong-Cheol, Lee Seoung Hoon, Kwon Kang-Beom, Lee Young-Rae
Center for Metabolic Function Regulation (CMFR), Wonkwang University School of Medicine, Iksan, 570-749, Republic of Korea.
Department of Rehabilitation Medicine of Korean Medicine, Wonkwang University School of Korean Medicine, Iksan City, Jeonbuk, 570-749, Republic of Korea.
BMC Complement Altern Med. 2017 Mar 23;17(1):164. doi: 10.1186/s12906-016-1550-x.
Sophorae Flos (SF) is a composite of flowers and buds of Styphnolobium japonicum (L.) Schott and has been used in traditional Korean and Chinese medicine for the treatment of hemostasis and inflammation. Previous studies reported that SF possesses anti-obesity properties, as well as anti-allergic, anti-proliferative, and anti-inflammatory activities. However, the effect of SF in bone resorption has not been studies. In this study, we examined the potential of SF extract (SFE) to inhibit receptor activator of NF-κB ligand (RANKL) -induced osteoclast differentiation in cultured mouse-derived bone marrow macrophages (BMMs).
BMMs, that act as osteoclast precursors, were cultured with M-CSF (50 ng/ml) and RANKL (100 ng/ml) for 4 days to generate osteoclasts. Osteoclast differentiation was measured by tartrate-resistant acidic phosphatase (TRAP) staining and the TRAP solution assay. Osteoclast differentiation marker genes were analyzed by the quantitative real-time polymerase chain reaction analysis. RANKLs signaling pathways were confirmed through western blotting.
SFE significantly decreased osteoclast differentiation in a dose-dependent manner. SFE inhibited RANKL-induced osteoclastogenesis by suppressing NF-κB activation. By contrast, SFE did not affect phospholipase C gamma 2 or subsequent cAMP response element binding activation. SFE inhibited the RANKL-induced expression of nuclear factor of activated T cells c1 (NFATc1).
SFE attenuated the RANKL-mediated induction of NF-κB through inhibition of IκBα phosphorylation, which contributed to inhibiting of RANKL-induced osteoclast differentiation through downregulation of NFATc1.
槐花（SF）是槐（Styphnolobium japonicum (L.) Schott）的花和花蕾的复合物，在传统的韩国和中国医学中用于治疗止血和炎症。先前的研究报道，槐花具有抗肥胖特性以及抗过敏、抗增殖和抗炎活性。然而，槐花在骨吸收方面的作用尚未得到研究。在本研究中，我们检测了槐花提取物（SFE）在体外培养的小鼠骨髓巨噬细胞（BMMs）中抑制核因子κB受体活化因子配体（RANKL）诱导的破骨细胞分化的潜力。
作为破骨细胞前体的BMMs与巨噬细胞集落刺激因子（M-CSF，50 ng/ml）和RANKL（100 ng/ml）一起培养4天以生成破骨细胞。通过抗酒石酸酸性磷酸酶（TRAP）染色和TRAP溶液测定法测量破骨细胞分化。通过定量实时聚合酶链反应分析破骨细胞分化标志物基因。通过蛋白质印迹法确认RANKL信号通路。
SFE以剂量依赖性方式显著降低破骨细胞分化。SFE通过抑制NF-κB活化来抑制RANKL诱导的破骨细胞生成。相比之下，SFE不影响磷脂酶Cγ2或随后的cAMP反应元件结合激活。SFE抑制RANKL诱导的活化T细胞核因子c1（NFATc1）的表达。
SFE通过抑制IκBα磷酸化减弱RANKL介导的NF-κB诱导，这有助于通过下调NFATc1来抑制RANKL诱导的破骨细胞分化。
