Department of Joint Surgery, The First Affiliated Hospital of Dalian Medical University, Dalian, Liaoning 116011, P.R. China.
Department of Joint Surgery and Sports Medicine, Tianjin Union Medical Center, Tianjin 300121, P.R. China.
Mol Med Rep. 2019 Jan;19(1):461-467. doi: 10.3892/mmr.2018.9690. Epub 2018 Nov 23.
Lonicera japonica has been used in traditional Chinese medicine as an important medicinal plant, with the ability to inhibit osteoclast development and bone loss. However, it is not clear which active ingredient exerts these effects. (R)‑dehydroxyabscisic alcohol β‑D‑apiofuranosyl‑(1ˮ→6')‑β‑D‑glucopyranoside (DAG) is an active constituent isolated from Lonicera japonica. In the present study, the ST2 bone marrow stromal cell line was treated by DAG at different concentrations and the osteoblastic differentiation was explored by ELISA assay, Von Kossa staining, Alizarin Red S staining, reverse transcription‑quantitative polymerase chain reaction and western blot analysis. The results revealed that DAG promoted osteoblastic differentiation, as evidenced by increasing mineralization and alkaline phosphatase (ALP) activity, as well as the expression of genes encoding bone differentiation markers, including Alp, osteopontin (Opn) and osteocalcin (Ocn). In addition, DAG upregulated the gene expression of bone morphogenetic protein (Bmp)‑2, Bmp4, Wnt family member (Wnt)‑1, Wnt3 and runt‑related transcription factor 2 (Runx2), as well as the protein expression of phosphorylated‑mothers against decapentaplegic homolog (Smad) 1, Smad5 Smad8, β‑catenin and Runx2 in ST2 cells. The osteogenic effects induced by DAG were attenuated by the BMP antagonist Noggin and the WNT signaling pathway inhibitor Dickkopf related protein‑1. The data indicated that DAG promoted the osteoblastic differentiation of ST2 cells, at least partially through regulating the BMP/WNT signaling pathways. This provides scientific rationale for the development of DAG as a treatment for bone loss‑associated diseases, such as osteoporosis.
金银花在传统中药中被用作一种重要的药用植物,具有抑制破骨细胞发育和骨质流失的能力。然而,尚不清楚哪种活性成分发挥了这些作用。(R)-去羟表雄酮-β-D-吡喃葡萄糖醛酸基-(1′→6′)-β-D-葡萄糖苷(DAG)是从金银花中分离得到的一种活性成分。本研究采用不同浓度的 DAG 处理 ST2 骨髓基质细胞系,通过 ELISA 检测、Von Kossa 染色、茜素红 S 染色、逆转录-定量聚合酶链反应和 Western blot 分析探讨其成骨分化作用。结果表明,DAG 可促进成骨分化,表现为矿化和碱性磷酸酶(ALP)活性增加,以及骨分化标志物编码基因的表达增加,包括 Alp、骨桥蛋白(Opn)和骨钙素(Ocn)。此外,DAG 上调了骨形态发生蛋白(Bmp)-2、Bmp4、Wnt 家族成员(Wnt)-1、Wnt3 和 Runt 相关转录因子 2(Runx2)的基因表达,以及磷酸化 Smad 1、Smad5、Smad8、β-连环蛋白和 Runx2 蛋白的表达。DAG 诱导的成骨作用被 BMP 拮抗剂 Noggin 和 WNT 信号通路抑制剂 Dickkopf 相关蛋白-1 减弱。这些数据表明,DAG 通过调节 BMP/WNT 信号通路促进 ST2 细胞的成骨分化。这为 DAG 作为治疗与骨质疏松症等骨丢失相关疾病的药物开发提供了科学依据。