BACKGROUND Osteoporosis is a worldwide severe bone disease. This study aimed to evaluate the effect of polyphyllin VII on the genesis of osteoclasts from bone marrow macrophages (BMMs) and its potentiality as a therapeutic drug for osteoporosis. METHODS BMMs were induced to differentiate into osteoclasts by RANKL and M-CSF. The cells were then treated with various concentrations of polyphyllin VII. Intracellular reactive oxygen species (ROS) measurement assay, resorption pit formation assay, tartrate-resistant acid phosphatase (TRAP) staining and TRAP activity assessment, cell viability assay, active GTPase pull-down assay, immunofluorescent staining, immunoblotting, and RT-PCR were performed. RESULTS RANKL + M-CSF significantly increased TRAP activity, number of osteoclasts, number and area of lacunae, intracellular content of ROS, protein levels of Nox1, TRAF6, c-Src and p-PI3K, as well as the content of activated GTP-Rac1, which were significantly blocked by polyphyllin VII in a concentration-dependent manner. CONCLUSION These findings suggested that polyphyllin VII inhibited differentiation of BMMs into osteoclasts through suppressing ROS synthesis, which was modulated by TRAF6-cSrc-PI3k signal transduction pathway including GTP-Rac1 and Nox1. Polyphyllin VII could be a therapeutic drug for osteoporosis.

中文翻译：

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通过抑制TRAF6 / c-Src / PI3K途径和ROS的产生，多叶连蛋白VII减弱了RANKL诱导的破骨细胞分化。

背景技术骨质疏松症是世界范围内的严重骨病。这项研究的目的是评估多糖7对骨髓巨噬细胞（BMM）破骨细胞的发生及其作为骨质疏松症治疗药物的潜力。方法RANKL和M-CSF诱导BMMs分化为破骨细胞。然后用各种浓度的多叶绿素VII处理细胞。细胞内活性氧（ROS）测量测定，重吸收坑形成测定，抗酒石酸酸性磷酸酶（TRAP）染色和TRAP活性评估，细胞活力测定，活性GTPase下拉测定，免疫荧光染色，免疫印迹和RT-PCR分别为执行。结果RANKL + M-CSF显着增加了TRAP活性，破骨细胞数量，腔隙的数量和面积，ROS的细胞内含量，Nophyl1，TRAF6，c-Src和p-PI3K的蛋白质水平，以及活化的GTP-Rac1的含量，均以浓度依赖的方式被叶绿素VII阻断。结论这些发现表明，多叶绿素VII通过抑制ROS合成来抑制BMMs向破骨细胞的分化，ROS的合成受TRAF6-cSrc-PI3k信号转导途径（包括GTP-Rac1和Nox1）的调节。Polyphyllin VII可能是骨质疏松症的治疗药物。它由包括GTP-Rac1和Nox1在内的TRAF6-cSrc-PI3k信号转导途径调节。Polyphyllin VII可能是骨质疏松症的治疗药物。它由包括GTP-Rac1和Nox1在内的TRAF6-cSrc-PI3k信号转导途径调节。Polyphyllin VII可能是骨质疏松症的治疗药物。