Sanguisorba officinalis L. (Rosaceae) is a perennial herbaceous plant and its roots have been used as an important traditional medicine for over 2000 years. Ziyuglycoside I (Ziyu), an active compound isolated from the roots of S. officinalis L., has shown biological effects such as anti-oxidant, antiviral, and antiwrinkle activities. This study aimed to elucidate the underlying mechanisms of action of Ziyu on cytotoxicity, migration, and differentiation of pre-osteoblasts. Herein, at concentrations ranging from 1 to 100 μM, Ziyu was not cytotoxic against pre-osteoblasts. Alkaline phosphatase activity assay and staining, and migration assay showed that Ziyu increased cell migration and promoted early osteoblast differentiation, followed by the enhancement of mineralized nodule formation in a dose-dependent manner, as indicated by Alizarin Red S staining. In addition, Ziyu increased the protein levels of runt-related transcription factor 2 (RUNX2) during osteoblast differentiation, whereas it did not affect the phosphorylation of Smad1/5/8 and GSK3b and expression of β-catenin. Ziyu also activated ERK1/2 and mitogen-activated protein kinase during osteoblast differentiation, and ERK1/2 inhibitor attenuated Ziyu-mediated RUNX2 expression and nuclear accumulation. Furthermore, Ziyu-mediated early and late osteoblast differentiation was significantly suppressed by the inhibition of ERK1/2, which was accompanied by attenuation in the mRNA levels of osteoblast-related genes including bone sialoprotein, osteopontin, and osteocalcin. Taken together, the findings of this study provide evidence that Ziyu promotes cell migration, osteoblast differentiation, and bone mineralization and suggest a potential role for Ziyu in the treatment of bone diseases.

中文翻译：

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Ziyuglycoside I 通过 ERK1/2 上调 RUNX2 促进成骨细胞分化和骨矿化

地榆L.（蔷薇科）是一种多年生草本植物，其根作为重要的传统药物已有2000多年的历史。Ziyuglycoside I (Ziyu)，一种从根部分离的活性化合物S. officinalisL.，已显示出生物效应，例如抗氧化、抗病毒和抗皱活性。本研究旨在阐明紫玉对前成骨细胞的细胞毒性、迁移和分化的潜在作用机制。在此，浓度范围为 1 至 100μM，紫玉对前成骨细胞没有细胞毒性。碱性磷酸酶活性测定和染色以及迁移测定显示紫玉增加细胞迁移并促进早期成骨细胞分化，随后以剂量依赖性方式增强矿化结节形成，如茜素红S染色所示。此外，紫玉在成骨细胞分化过程中增加了runt相关转录因子2（RUNX2）的蛋白水平，但不影响Smad1/5/8和GSK3b的磷酸化和β-连环蛋白。Ziyu 还在成骨细胞分化过程中激活 ERK1/2 和丝裂原活化蛋白激酶，ERK1/2 抑制剂减弱 Ziyu 介导的 RUNX2 表达和核积累。此外，子玉介导的早期和晚期成骨细胞分化受到抑制 ERK1/2 的显着抑制，伴随着成骨细胞相关基因（包括骨唾液酸蛋白、骨桥蛋白和骨钙素）的 mRNA 水平减弱。综上所述，本研究的结果提供了紫玉促进细胞迁移、成骨细胞分化和骨矿化的证据，并表明紫玉在治疗骨疾病中的潜在作用。