Nagaoka Masahiro, Maeda Toyonobu, Chatani Masahiro, Handa Kazuaki, Yamakawa Tomoyuki, Kiyohara Shuichi, Negishi-Koga Takako, Kato Yasumasa, Takami Masamichi, Niida Shumpei, Lang Stefanie C, Kruger Marlena C, Suzuki Keiko
Department of Pharmacology, School of Dentistry, Ohu University, Fukushima 963-8611, Japan.
Department of Oral Function and Molecular Biology, School of Dentistry, Ohu University, Fukushima 963-8611, Japan.
Antioxidants (Basel). 2019 Sep 10;8(9):386. doi: 10.3390/antiox8090386.
In our previous investigation, delphinidin, one of the most abundant anthocyanins found in vegetables and berry fruits, had been shown to inhibit osteoclasts and prevent bone loss in mouse models of osteoporosis. In the present study, we investigated whether a delphinidin glycoside-enriched maqui berry extract (MBE, Delphinol) exhibits beneficial effects on bone metabolism both in vitro and in vivo. MBE stimulated the osteoblastic differentiation of MC3T3-E1 cells, as indicated by enhanced mineralized nodule formation, and increased alkaline phosphatase activity, through the upregulation of bone morphogenetic protein 2 (), runt-related transcription factor 2 (), Osterix (), osteocalcin (), and matrix extracellular phosphoglycoprotein () mRNA expression. Immunostaining and immunoprecipitation assays demonstrated that MBE suppressed NF-κB transnucleation through acting as a superoxide anion/peroxynitrite scavenger in MC3T3-E1 cells. Simultaneously, MBE inhibited both osteoclastogenesis in primary bone marrow macrophages and pit formation by maturated osteoclasts on dentine slices. Microcomputed tomography (micro-CT) and bone histomorphometry analyses of femurs demonstrated that the daily ingestion of MBE significantly increased BV/TV (ratio of bone volume to tissue volume), Tb.Th (trabecular thickness), Tb.N (trabecular number), N.Nd/N.Tm (node to terminus ratio), OV/TV (ratio of osteoid volume to tissue volume), BFR/TV (bone formation rate per tissue volume), and significantly decreased Tb.Sp (trabecular separation), ES/BS (ratio of eroded surface to bone surface) and N.Oc/BS (number of osteoclast per unit of bone surface, compared to vehicle controls in osteopenic mouse models. These findings suggest that MBE can be a promising natural agent for the prevention of bone loss in osteopenic conditions by not only inhibiting bone resorption, but also stimulating bone formation.
在我们之前的研究中，飞燕草素是蔬菜和浆果中含量最为丰富的花青素之一，已被证实在骨质疏松症小鼠模型中可抑制破骨细胞并预防骨质流失。在本研究中，我们调查了富含飞燕草素糖苷的智利酒果提取物（MBE，Delphinol）在体外和体内对骨代谢是否具有有益作用。MBE通过上调骨形态发生蛋白2（）、 runt相关转录因子2（）、Osterix（）、骨钙素（）和基质细胞外磷酸糖蛋白（）的mRNA表达，刺激MC3T3-E1细胞的成骨细胞分化，这表现为矿化结节形成增加以及碱性磷酸酶活性增强。免疫染色和免疫沉淀分析表明，MBE在MC3T3-E1细胞中作为超氧阴离子/过氧亚硝酸盐清除剂发挥作用，从而抑制NF-κB转核。同时，MBE抑制原代骨髓巨噬细胞中的破骨细胞生成以及成熟破骨细胞在牙本质切片上形成的陷窝。对股骨进行的微型计算机断层扫描（micro-CT）和骨组织形态计量学分析表明，每日摄入MBE可显著增加骨体积与组织体积之比（BV/TV）、骨小梁厚度（Tb.Th）、骨小梁数量（Tb.N）、节点与末端比率（N.Nd/N.Tm）、类骨质体积与组织体积之比（OV/TV）、每组织体积骨形成率（BFR/TV），并显著降低骨小梁间距（Tb.Sp）、侵蚀表面与骨表面之比（ES/BS）以及每单位骨表面破骨细胞数量（N.Oc/BS），与骨质疏松症小鼠模型中的赋形剂对照组相比。这些发现表明，MBE不仅可以抑制骨吸收，还可以刺激骨形成，有望成为预防骨质疏松症骨质流失的天然药物。
