PDF(Pubmed)
Abstract：
UNASSIGNED: Nitrate, a key component of saliva, has been shown widely physiological functions in the human body. But its function on bone metabolism remains unclear. The aim of this study was to investigate the function and mechanism of saliva nitrate on osteoporosis and the function of bone marrow mesenchymal stem cells (BMSCs).
UNASSIGNED: Saliva nitrate removal or supplemental interventions were performed for 1 month in ovariectomized (OVX) osteopenia mice. The nitrate levels in saliva and serum were detected. The bone formation and bone microarchitecture in the OVX mouse model were investigated by quantitative Micro--computed tomography imaging, histological staining and serum bone biomarker analysis. The effects of nitrate on the functional homeostasis of BMSCs in OVX mice were explored by Ki67 immunofluorescence staining, Ki67 flow staining, alizarin red staining, qPCR and western blotting. Finally, downstream signaling pathways were screened by proteomics and verified by western blotting.
UNASSIGNED: The results showed that nitrate deficiency exacerbated osteoporosis, while nitrate administration prevent osteoporosis in OVX mice. Nitrate increased the expression of PINP, a biomarker of bone formation, in OVX mice. Besides, nitrate enhanced the proliferative capacity and osteogenic function of BMSCs in OVX mice in vitro and in vivo. In addition, nitrate upregulated the expression levels of osteogenesis-related genes ALP, Run2 and OPN of BMSCs. EGFR and mTOR signaling were screened as the key downstream of nitrate, and phosphorylated protein levels of its subfamily members AKT, ERK and S6K were significantly upregulated by nitrate.
UNASSIGNED: The present results showed saliva nitrate preventively protects against osteoporosis through enhances the proliferation and osteogenic differentiation potential of BMSCs. The effects of nitrate on bone homeostasis are closely related to the EGFR/AKT/ERK and mTOR/S6K signaling axes.
UNASSIGNED: Our study provides experimental evidence for the use of saliva nitrate as an effective candidate for the prevention of osteoporosis and maintenance of bone homeostasis.
UNASSIGNED: Saliva nitrate removal or supplemental interventions were performed for 1 month in ovariectomized (OVX) osteopenia mice. The nitrate levels in saliva and serum were detected. The bone formation and bone microarchitecture in the OVX mouse model were investigated by quantitative Micro--computed tomography imaging, histological staining and serum bone biomarker analysis. The effects of nitrate on the functional homeostasis of BMSCs in OVX mice were explored by Ki67 immunofluorescence staining, Ki67 flow staining, alizarin red staining, qPCR and western blotting. Finally, downstream signaling pathways were screened by proteomics and verified by western blotting.
UNASSIGNED: The results showed that nitrate deficiency exacerbated osteoporosis, while nitrate administration prevent osteoporosis in OVX mice. Nitrate increased the expression of PINP, a biomarker of bone formation, in OVX mice. Besides, nitrate enhanced the proliferative capacity and osteogenic function of BMSCs in OVX mice in vitro and in vivo. In addition, nitrate upregulated the expression levels of osteogenesis-related genes ALP, Run2 and OPN of BMSCs. EGFR and mTOR signaling were screened as the key downstream of nitrate, and phosphorylated protein levels of its subfamily members AKT, ERK and S6K were significantly upregulated by nitrate.
UNASSIGNED: The present results showed saliva nitrate preventively protects against osteoporosis through enhances the proliferation and osteogenic differentiation potential of BMSCs. The effects of nitrate on bone homeostasis are closely related to the EGFR/AKT/ERK and mTOR/S6K signaling axes.
UNASSIGNED: Our study provides experimental evidence for the use of saliva nitrate as an effective candidate for the prevention of osteoporosis and maintenance of bone homeostasis.
摘要：
■硝酸盐，唾液的关键成分，已经在人体中显示出广泛的生理功能。但其对骨代谢的作用尚不清楚。本研究旨在探讨唾液硝酸盐对骨质疏松的作用机制及骨髓间充质干细胞(BMSCs)的作用。
■在卵巢切除(OVX)骨质减少小鼠中进行唾液硝酸盐去除或补充干预1个月。检测唾液和血清中的硝酸盐含量。通过定量显微计算机断层扫描成像研究OVX小鼠模型中的骨形成和骨微结构，组织学染色和血清骨生物标志物分析。通过Ki67免疫荧光染色探讨硝酸盐对OVX小鼠BMSCs功能稳态的影响,Ki67流染色，茜素红染色，qPCR和蛋白质印迹。最后,通过蛋白质组学筛选下游信号通路,并通过蛋白质印迹进行验证.
■结果显示硝酸盐缺乏会加剧骨质疏松,而硝酸盐给药可预防OVX小鼠的骨质疏松症。硝酸盐增加PINP的表达，骨形成的生物标志物，在OVX小鼠中。此外,硝酸盐在体外和体内增强了OVX小鼠BMSCs的增殖能力和成骨功能。此外,硝酸盐上调成骨相关基因ALP的表达水平,BMSCs的Run2和OPN。EGFR和mTOR信号被筛选为硝酸盐的关键下游，及其亚家族成员AKT的磷酸化蛋白水平，ERK和S6K被硝酸盐显著上调。
■目前的结果表明，唾液硝酸盐通过增强BMSCs的增殖和成骨分化潜能来预防性预防骨质疏松症。硝酸盐对骨稳态的影响与EGFR/AKT/ERK和mTOR/S6K信号轴密切相关。
■我们的研究为使用唾液硝酸盐作为预防骨质疏松症和维持骨稳态的有效候选者提供了实验证据。
■在卵巢切除(OVX)骨质减少小鼠中进行唾液硝酸盐去除或补充干预1个月。检测唾液和血清中的硝酸盐含量。通过定量显微计算机断层扫描成像研究OVX小鼠模型中的骨形成和骨微结构，组织学染色和血清骨生物标志物分析。通过Ki67免疫荧光染色探讨硝酸盐对OVX小鼠BMSCs功能稳态的影响,Ki67流染色，茜素红染色，qPCR和蛋白质印迹。最后,通过蛋白质组学筛选下游信号通路,并通过蛋白质印迹进行验证.
■结果显示硝酸盐缺乏会加剧骨质疏松,而硝酸盐给药可预防OVX小鼠的骨质疏松症。硝酸盐增加PINP的表达，骨形成的生物标志物，在OVX小鼠中。此外,硝酸盐在体外和体内增强了OVX小鼠BMSCs的增殖能力和成骨功能。此外,硝酸盐上调成骨相关基因ALP的表达水平,BMSCs的Run2和OPN。EGFR和mTOR信号被筛选为硝酸盐的关键下游，及其亚家族成员AKT的磷酸化蛋白水平，ERK和S6K被硝酸盐显著上调。
■目前的结果表明，唾液硝酸盐通过增强BMSCs的增殖和成骨分化潜能来预防性预防骨质疏松症。硝酸盐对骨稳态的影响与EGFR/AKT/ERK和mTOR/S6K信号轴密切相关。
■我们的研究为使用唾液硝酸盐作为预防骨质疏松症和维持骨稳态的有效候选者提供了实验证据。
