Icariside II promotes the osteogenic differentiation of canine bone marrow mesenchymal stem cells via the PI3K/AKT/mTOR/S6K1 signaling pathways.
Icariside II promotes the osteogenic differentiation of canine bone marrow mesenchymal stem cells via the PI3K/AKT/mTOR/S6K1 signaling pathways.
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发表时间：
2017
影响因子：
2.2
通讯作者：
Guangming Luo;Biao Xu;Yuanliang Huang
中科院分区：
文献类型：
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作者：
Guangming Luo;Biao Xu;Yuanliang Huang
本研究旨在探讨淫羊藿苷II（ICSII）对犬骨髓间充质干细胞（BMSCs）的成骨作用及其诱导途径。培养扩增至第4代。使用细胞计数试剂盒-8（CCK-8）测定来评估ICSII的增殖作用。碱性磷酸酶（ALP）活性测定和茜素红染色检测ICSII对BMSCs体外成骨的影响。采用Western blot和实时荧光定量PCR（RT-PCR）检测成骨蛋白/基因的表达。我们还评估了Akt和S6 K1磷酸化水平的变化，沿着在ICSII存在或不存在下用渥曼青霉素（磷脂酰肌醇3-激酶抑制剂; PI 3 K）或雷帕霉素（哺乳动物雷帕霉素靶蛋白（mTOR）的特异性抑制剂）预处理后成骨蛋白/基因表达的变化。我们的研究结果表明，ICSII促进BMSCs的增殖，同时抑制ALP活性。我们还发现BMSC用ICSII和成骨培养基处理21天后形成钙结节。ICSII显著增加成骨蛋白/基因的表达，并提高Akt和S6 K1的磷酸化水平。用渥曼青霉素或雷帕霉素处理减弱了p-Akt、p-S6 K1和成骨蛋白/基因的表达。这些结果表明ICSII通过PI 3 K/AKT/mTOR/S6 K1信号通路促进犬BMSCs的成骨分化。
The aim of the present study was to investigate the osteogenic effects of icariside II (ICSII) on canine bone marrow mesenchymal stem cells (BMSCs) and the pathways by which these effects were induced. BMSCs were cultured and expanded to the fourth passage. The proliferative effects of ICSII were assessed using the cell counting kit-8 (CCK-8) assay. The osteogenic response to ICSII in BMSCs in vitro was examined by alkaline phosphatase (ALP) activity assays and Alizarin red staining. Using Western blot assays and real-time PCR (RT-PCR), we examined the expression of osteogenetic proteins/genes. We also evaluated changes in Akt and S6K1 phosphorylation levels, along with changes in the expression of osteogenesis proteins/genes after pretreatment with wortmannin (an inhibitor of phosphatidylinositol 3-kinase; PI3K) or rapamycin [a specific inhibitor of mammalian target of rapamycin (mTOR)] in the presence or absence of ICSII. Our results show that ICSII promotes the proliferation of BMSCs, while inhibiting ALP activity. We also found that calcium nodules form after BMSCs are treated with ICSII and osteogenetic medium for 21 days. ICSII significantly increased the expression of osteogenesis proteins/genes and elevated the phosphorylation levels of Akt and S6K1. Treatment with wortmannin or rapamycin attenuated the expression of p-Akt, p-S6K1, and osteogenesis proteins/genes. These results suggest that ICSII promotes the osteogenic differentiation of canine BMSCs via the PI3K/AKT/mTOR/S6K1 signaling pathways.