Combined Effect of Midazolam and Bone Morphogenetic Protein-2 for Differentiation Induction from C2C12 Myoblast Cells to Osteoblasts
Combined Effect of Midazolam and Bone Morphogenetic Protein-2 for Differentiation Induction from C2C12 Myoblast Cells to Osteoblasts
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DOI：
10.3390/pharmaceutics12030218
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发表时间：
2020-03-01
期刊：
影响因子：
5.4
通讯作者：
Kawahara, Hiroshi
中科院分区：
文献类型：
--
作者：
Hidaka, Yukihiko;Chiba-Ohkuma, Risako;Kawahara, Hiroshi
在药物重新定位研究中，为现有药物确定了药物发现的新概念和新的治疗机会。咪达唑仑(MDZ)是一种用于全身麻醉的麻醉诱导剂。在这里，我们展示了骨形态发生蛋白-2(BMP-2)和MDZ在成骨分化中的联合作用。用BMP-2和MDZ(BMP-2+MDZ)联合培养永生化小鼠成肌细胞系C2C12细胞。从生物学、免疫组织化学和遗传细胞水平研究了C2C12细胞向成骨细胞的分化和信号转导。从晶体工程水平对C2C12细胞形成的矿化结节进行了表征。BMP-2+MDZ处理使C2C12细胞肌管形成减少，碱性磷酸酶活性增强，成骨细胞分化标志基因表达水平升高。析出的结节由随机取向的羟基磷灰石纳米棒和纳米颗粒组成。BMP-2+MDZ处理降低了C2C12细胞GABAA受体上α1和γ2亚单位抗原的免疫染色，但增强了BMP信号转导的免疫染色。我们的研究表明，BMP-2+MDZ具有较强的诱导C2C12细胞向成骨细胞分化的能力，在骨再生中具有药物重定位的潜力。
In drug repositioning research, a new concept in drug discovery and new therapeutic opportunities have been identified for existing drugs. Midazolam (MDZ) is an anesthetic inducer used for general anesthesia. Here, we demonstrate the combined effects of bone morphogenetic protein-2 (BMP-2) and MDZ on osteogenic differentiation. An immortalized mouse myoblast cell line (C2C12 cell) was cultured in the combination of BMP-2 and MDZ (BMP-2+MDZ). The differentiation and signal transduction of C2C12 cells into osteoblasts were investigated at biological, immunohistochemical, and genetic cell levels. Mineralized nodules formed in C2C12 cells were characterized at the crystal engineering level. BMP-2+MDZ treatment decreased the myotube cell formation of C2C12 cells, and enhanced alkaline phosphatase activity and expression levels of osteoblastic differentiation marker genes. The precipitated nodules consisted of randomly oriented hydroxyapatite nanorods and nanoparticles. BMP-2+MDZ treatment reduced the immunostaining for both alpha 1 and gamma 2 subunits antigens on the gamma-aminobutyric acid type A (GABAA) receptor in C2C12 cells, but enhanced that for BMP signal transducers. Our investigation showed that BMP-2+MDZ has a strong ability to induce the differentiation of C2C12 cells into osteoblasts and has the potential for drug repositioning in bone regeneration.