Biological Sciences Department, College of Science, King Faisal University, Hofuf-31982, Al-Ahsa, Saudi Arabia.
Endocrine Research (KMEB), Department of Endocrinology, Odense University Hospital and University of Southern Denmark, Odense, Denmark.
J Biomed Sci. 2019 Jul 5;26(1):51. doi: 10.1186/s12929-019-0544-7.
Identifying bone anabolic agents is a superior strategy for the treatment of osteoporosis. Naturally, derived coumarin derivatives have shown osteoanabolic effect in vitro and in vivo. In this study, we investigated the effect of 5'-Hydroxy Auraptene (5'-HA), a coumarin derivative that newly isolated from Lotus lalambensis Schweinf on the differentiation of the mouse bone marrow-derived mesenchymal (skeletal) stem cells (mBMSCs) into osteoblast and adipocyte.
The effect of 5'-HA on mBMSCs cell proliferation and osteoblast differentiation was assessed by measuring cell viability, quantitative alkaline phosphatase (ALP) activity assay, Alizarin red staining for matrix mineralization and osteogenic gene array expression. Adipogenesis was measured by Oil Red O staining and quantitative real time PCR (qPCR) analysis of adipogenic markers. Regulation of BMPs signaling pathways by 5'-HA was measured by Western blot analysis and qPCR.
5'-HA showed to stimulate the differentiation of mBMSCs into osteogenic cell lineage in a dose-dependent manner, without affecting their differentiation into adipocytic cell lineage. Treatment of mBMSCs with 5'-HA showed to promote significantly the BMP2-induced osteogenesis in mBMSCs via activating Smad1/5/8 phosphorylation and increasing Smad4 expression. Blocking of BMP signaling using BMPR1 selective inhibitor LDN-193189 significantly inhibited the stimulatory effect of 5'-HA on osteogenesis.
Our data identified 5'-HA, as a novel coumarin derivative that function to stimulate the differentiation of mBMSCs into osteoblasts in BMP-signaling dependent mechanism.
鉴定骨合成代谢药物是治疗骨质疏松症的一种更好的策略。天然衍生的香豆素衍生物已在体外和体内显示出成骨作用。在这项研究中，我们研究了 5'-羟基花椒毒素（5'-HA），一种新从 Lotus lalambensis Schweinf 中分离出的香豆素衍生物，对小鼠骨髓间充质（骨骼）干细胞（mBMSCs）向成骨细胞和脂肪细胞分化的影响。
通过测量细胞活力、定量碱性磷酸酶（ALP）活性测定、基质矿化的茜素红染色和成骨基因表达谱，评估 5'-HA 对 mBMSCs 细胞增殖和成骨分化的影响。通过油红 O 染色和脂肪生成标志物的定量实时 PCR（qPCR）分析来测量脂肪生成。通过 Western blot 分析和 qPCR 来测量 5'-HA 对 BMPs 信号通路的调节作用。
5'-HA 以剂量依赖的方式刺激 mBMSCs 向成骨细胞谱系分化，而不影响其向脂肪细胞谱系分化。用 5'-HA 处理 mBMSCs 可通过激活 Smad1/5/8 磷酸化和增加 Smad4 表达，显著促进 BMP2 诱导的 mBMSCs 成骨作用。使用 BMPR1 选择性抑制剂 LDN-193189 阻断 BMP 信号显著抑制了 5'-HA 对成骨作用的刺激作用。
我们的数据确定 5'-HA 是一种新型香豆素衍生物，通过 BMP 信号依赖性机制刺激 mBMSCs 向成骨细胞分化。
