BACKGROUND Astragalus saponin IV(AS- IV) extracted from tranditional Chinese medicine Radix Astragali Mongolici, which had been reported to have medicinal properties in treating several types of diseases. This study aimed at investigating the biological functions of AS-IV on bone marrow mesenchymal cells(BMSCs) differentiation, therefore, seeking for a better application of AS-IV on fracture or other orthopedic disorders. METHODS AS-IV was co-incubated with BMSCs in vitro to testify whether it can influence the proliferation and differentiation of BMSCs. Cell proliferation activity was detected by Cell Counting Kit-8 (CCK-8), while its differentiation promoting capibility was obtained by alkaline phosphatase (ALP) activity assay and Alizarin red S staining. Besides, differentiation protein markers of preosteoblast was detected by western blots. Neuron growth factor antagonists (NGFA) and microRNA-21 (miR-21) inhibitors were co incubated with AS-IV to search the regulatory pathways it activated in BMSCs. RESULTS AS-IV incubation boosted the proliferation of BMSCs, and accelerated the differentiating direction into preosteoblasts. Runx2, OPN, BMP2, OCN proteins were up regulated after AS- IV treatment. MiR-21/NGF/BMP2/Runx2 pathway can participate the biological effects of AS- IV on BMSCs. CONCLUSION AS- IV might be used as a therapeutic agent for bone fracture or other orthopedic disorders.

中文翻译：

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黄芪皂苷IV通过miR-21 / NGF / BMP2 / Runx2途径促进骨髓间充质干细胞的成骨分化。

背景技术从传统中药黄芪中提取的黄芪皂苷IV（AS-IV），据报道具有治疗多种类型疾病的药用特性。这项研究旨在调查AS-IV对骨髓间充质细胞（BMSCs）分化的生物学功能，因此，寻求AS-IV在骨折或其他骨科疾病中的更好应用。方法将AS-IV与骨髓间充质干细胞在体外共同孵育，以验证其是否能影响骨髓间充质干细胞的增殖和分化。细胞增殖活性通过Cell Counting Kit-8（CCK-8）检测，而其分化促进能力则通过碱性磷酸酶（ALP）活性测定和茜素红S染色获得。此外，通过western印迹检测了成骨细胞的分化蛋白标记。将神经元生长因子拮抗剂（NGFA）和microRNA-21（miR-21）抑制剂与AS-IV共同孵育，以研究其在BMSC中激活的调节途径。结果AS-IV温育促进了BMSCs的增殖，并加速了分化为成骨细胞的方向。AS-IV处理后，Runx2，OPN，BMP2，OCN蛋白被上调。MiR-21 / NGF / BMP2 / Runx2途径可以参与AS-IV对BMSC的生物学作用。结论AS-IV可以用作骨折或其他骨科疾病的治疗剂。AS-IV处理后，BMP2，OCN蛋白被上调。MiR-21 / NGF / BMP2 / Runx2途径可以参与AS-IV对BMSC的生物学作用。结论AS-IV可用作骨折或其他骨科疾病的治疗剂。AS-IV处理后，BMP2，OCN蛋白上调。MiR-21 / NGF / BMP2 / Runx2途径可以参与AS-IV对BMSC的生物学作用。结论AS-IV可以用作骨折或其他骨科疾病的治疗剂。