Department of Oral and Maxillofacial Pathology, School of Dentistry, Kyung Hee University, Seoul 02447, Republic of Korea.
National Development Institute for Korean Medicine, Gyeongsan 38540, Republic of Korea.
Cells. 2022 Dec 1;11(23):3887. doi: 10.3390/cells11233887.
Gentianae Scabrae Radix is used in traditional medicine and is known to possess bioactive compounds, including secoiridoid glycosides, flavonoids, lignans, and triterpenes. Trifloroside (TriFs) is a secoiridoid glycoside known for its antioxidant activity; however, its other effects have not been studied. In the present study, we investigated the biological effects of TriFs isolated from the roots of Gentianae Scabrae Radix using pre-osteoblast MC3T3E-1 cells. No cellular toxicity was observed with 1 μM TriFs, whereas 5-100 μM TriFs showed a gradual increase in cell viability. Alkaline phosphatase staining and microscopic observations revealed that 1-10 μM TriFs stimulated osteogenic activity during early osteoblast differentiation. Trifloroside also increased mineral apposition during osteoblast maturation. Biochemical analyses revealed that TriFs promoted nuclear RUNX2 expression and localization by stimulating the major osteogenic BMP2-Smad1/5/8-RUNX2 pathway. Trifloroside also increased p-GSK3β, β-catenin, p-JNK, and p-p38, but not Wnt3a, p-AKT, and p-ERK. Moreover, TriFs increased the MMP13 levels and promoted cell migration and adhesion. In contrast, TriFs-induced osteoblast differentiation and maturation had negligible effects on autophagy and necrosis. Our findings suggest that TriFs induces osteogenic effects through differentiation, adhesion, migration, and mineral apposition. Therefore, TriFs is suggested as a potential drug target in osteoblast-mediated bone diseases.
獐牙菜苦苷是一种传统中药，已知含有生物活性化合物，包括裂环环烯醚萜苷类、黄酮类、木脂素和三萜类。獐牙菜苦苷（TriFs）是一种裂环环烯醚萜苷类，具有抗氧化活性；然而，其其他作用尚未得到研究。在本研究中，我们使用前成骨细胞 MC3T3E-1 细胞研究了从獐牙菜苦苷根部分离得到的 TriFs 的生物学效应。1 μM TriFs 无细胞毒性，而 5-100 μM TriFs 则表现出细胞活力逐渐增加。碱性磷酸酶染色和显微镜观察显示，1-10 μM TriFs 刺激早期成骨细胞分化过程中的成骨活性。獐牙菜苦苷还通过刺激主要成骨 BMP2-Smad1/5/8-RUNX2 途径促进成骨细胞成熟过程中的矿化。TriFs 还通过刺激主要成骨 BMP2-Smad1/5/8-RUNX2 途径促进核 RUNX2 表达和定位。TriFs 还增加了 p-GSK3β、β-catenin、p-JNK 和 p-p38，但不增加 Wnt3a、p-AKT 和 p-ERK。此外，TriFs 增加了 MMP13 水平并促进了细胞迁移和黏附。相反，TriFs 诱导的成骨细胞分化和成熟对自噬和坏死几乎没有影响。我们的研究结果表明，TriFs 通过分化、黏附、迁移和矿化诱导成骨作用。因此，TriFs 被认为是成骨细胞介导的骨疾病的潜在药物靶点。
