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柚皮苷通过激活人骨髓间充质干细胞中的ERK信号通路增强成骨分化。

Naringin enhances osteogenic differentiation through the activation of ERK signaling in human bone marrow mesenchymal stem cells.

作者信息

Wang Huichao, Li Chunbo, Li Jianming, Zhu Yingjie, Jia Yudong, Zhang Ying, Zhang Xiaodong, Li Wenlong, Cui Lei, Li Wuyin, Liu Youwen

机构信息

Luoyang Orthopedic Hospital of Henan Province, Orthopedic Institute of Henan Province, Luoyang 471002, China.

Shanghai First Maternity and Infant Hospital, Tongji University of Medicine. Shanghai, 200126, China.

出版信息

Iran J Basic Med Sci. 2017 Apr;20(4):408-414. doi: 10.22038/IJBMS.2017.8582.

Abstract

OBJECTIVES

Naringin has been reported to regulate bone metabolism. However, its effect on osteogenesis remains unclear. The aim was to investigate the effect of naringin on osteogenic differentiation of human bone marrow mesenchymal stem cells (hBMSCs) through the activation of the ERK signaling pathway in osteogenic differentiation.

MATERIALS AND METHODS

Annexin V-FITC assay and MTT assay were used to measure the effect of naringin on cytotoxicity and proliferation of hBMSCs, respectively. Alkaline phosphatase activity analysis, Alizarin Red S staining, Western blotting, and real-time PCR assay were used to evaluate both the potential effect of naringin on osteogenic differentiation and the role of ERK signaling pathway in osteogenic differentiation.

RESULTS

Our results showed that naringin had no obvious toxicity on hBMSCs, and could significantly promote the proliferation of hBMSCs. Naringin also enhanced the osteogenic differentiation of hBMSCs and increased the protein and mRNA expression levels of osteogenic markers such as Runx-2, OXS, OCN, and Col1 in a dose-dependent manner. In addition, we found that the enhancing effect of naringin on osteogenic differentiation was related to the activation of phosphor-ERK, with an increase in duration of activity from 30 min to 120 min. More importantly, both the enhancing effect of naringin on osteogenic differentiation and the activity effect of naringin on ERK signaling pathway were reversed by U0126 addition.

CONCLUSION

Our findings demonstrated that naringin promoted proliferation and osteogenesis of hBMSCs by activating the ERK signaling pathway and it might be a potential therapeutic agent for treating or preventing osteoporosis.

摘要

目的

据报道,柚皮苷可调节骨代谢。然而,其对成骨作用的影响仍不清楚。本研究旨在探讨柚皮苷通过激活成骨分化过程中的ERK信号通路,对人骨髓间充质干细胞(hBMSCs)成骨分化的影响。

材料与方法

分别采用Annexin V-FITC法和MTT法检测柚皮苷对hBMSCs细胞毒性和增殖的影响。采用碱性磷酸酶活性分析、茜素红S染色、蛋白质印迹法和实时PCR检测法,评估柚皮苷对成骨分化的潜在作用以及ERK信号通路在成骨分化中的作用。

结果

结果显示,柚皮苷对hBMSCs无明显毒性,且能显著促进hBMSCs的增殖。柚皮苷还能增强hBMSCs的成骨分化,并以剂量依赖的方式增加成骨标志物如Runx-2、OXS、OCN和Col1的蛋白质和mRNA表达水平。此外,我们发现柚皮苷对成骨分化的增强作用与磷酸化ERK的激活有关,活性持续时间从30分钟增加到120分钟。更重要的是,添加U0126可逆转柚皮苷对成骨分化的增强作用以及对ERK信号通路的激活作用。

结论

我们的研究结果表明,柚皮苷通过激活ERK信号通路促进hBMSCs的增殖和成骨,可能是治疗或预防骨质疏松症的潜在治疗药物。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f928/5425923/eeac78260d56/IJBMS-20-408-g001.jpg

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