Department of Oral and Maxillofacial Pathology, School of Dentistry, Kyung Hee University, Seoul 02447, Korea.
National Institute for Korean Medicine Development, Gyeongsan 38540, Korea.
Int J Mol Sci. 2020 Dec 16;21(24):9579. doi: 10.3390/ijms21249579.
Sieb. et Zucc. has been used as traditional medicine in inflammatory diseases, and isolated compounds have shown pharmacological activities. Pinoresinol glucoside (PIN) belonging to lignins was isolated from the stem bark of This study aimed to investigate the biological function and mechanisms of PIN on cell migration, osteoblast differentiation, and matrix mineralization. Herein, we investigated the effects of PIN in MC3T3-E1 pre-osteoblasts, which are widely used for studying osteoblast behavior in in vitro cell systems. At concentrations ranging from 0.1 to 100 μM, PIN had no cell toxicity in pre-osteoblasts. Pre-osteoblasts induced osteoblast differentiation, and the treatment of PIN (10 and 30 μM) promoted the cell migration rate in a dose-dependent manner. At concentrations of 10 and 30 μM, PIN elevated early osteoblast differentiation in a dose-dependent manner, as indicated by increases in alkaline phosphatase (ALP) staining and activity. Subsequently, PIN also increased the formation of mineralized nodules in a dose-dependent manner, as indicated by alizarin red S (ARS) staining, demonstrating positive effects of PIN on late osteoblast differentiation. In addition, PIN induced the mRNA level of BMP2, ALP, and osteocalcin (OCN). PIN also upregulated the protein level of BMP2 and increased canonical BMP2 signaling molecules, the phosphorylation of Smad1/5/8, and the protein level of Runt-related transcription factor 2 (RUNX2). Furthermore, PIN activated non-canonical BMP2 signaling molecules, activated MAP kinases, and increased β-catenin signaling. The findings of this study indicate that PIN has biological roles in osteoblast differentiation and matrix mineralization, and suggest that PIN might have anabolic effects in bone diseases such as osteoporosis and periodontitis.
Siebold 和 Zucc. 已被用作治疗炎症性疾病的传统药物，其分离得到的化合物具有药理学活性。木脂素类的松柏脂素葡萄糖苷（PIN）从该植物的茎皮中分离得到。本研究旨在探讨 PIN 对细胞迁移、成骨细胞分化和基质矿化的生物学功能和作用机制。在此，我们研究了 PIN 对 MC3T3-E1 前成骨细胞的影响，该细胞广泛用于研究体外细胞系统中成骨细胞的行为。在 0.1 至 100 μM 的浓度范围内，PIN 对前成骨细胞没有细胞毒性。前成骨细胞诱导成骨细胞分化，PIN（10 和 30 μM）处理以剂量依赖性方式促进细胞迁移率。在 10 和 30 μM 浓度下，PIN 以剂量依赖性方式增加早期成骨细胞分化，碱性磷酸酶（ALP）染色和活性增加表明这一点。随后，PIN 还以剂量依赖性方式增加矿化结节的形成，茜素红 S（ARS）染色表明这一点，表明 PIN 对晚期成骨细胞分化有积极作用。此外，PIN 诱导 BMP2、ALP 和骨钙素（OCN）的 mRNA 水平增加。PIN 还上调 BMP2 蛋白水平，并增加经典 BMP2 信号分子，Smad1/5/8 的磷酸化和 Runt 相关转录因子 2（RUNX2）的蛋白水平。此外，PIN 激活非经典 BMP2 信号分子，激活 MAP 激酶，并增加 β-连环蛋白信号。本研究的结果表明，PIN 在成骨细胞分化和基质矿化中具有生物学作用，并表明 PIN 可能在骨质疏松症和牙周炎等骨病中具有合成代谢作用。
