Naringenin promotes SDF-1/CXCR4 signaling pathway in BMSCs osteogenic differentiation.

IF 16.4 1区化学 Q1 CHEMISTRY, MULTIDISCIPLINARY

Accounts of Chemical Research Pub Date : 2021-01-01 Epub Date: 2021-03-11 DOI:10.5603/FHC.a2021.0008

Yipei Wang, Shulin Bai, Qian Cheng, Yang Zeng, Xiaomei Xu, Guangzhao Guan

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引用次数: 0

Abstract

Introduction: Naringenin, a dihydro-flavonoid compound that shows chemotactic activity, may have a good application prospect in repairing bone tissue, but its specific mechanism in bone regeneration, especially the osteogenic differentiation of stem cells, needs for a further study. The aim of this study was to investigate the effect of naringenin on the osteogenic differentiation and its roles in the C-X-C chemokine receptor type 4/stromal cell-derived factor 1 (SDF-1/CXCR4) signal pathway of bone marrow-derived mesenchymal stem cells (BMSCs).

Material and methods: BMSCs were harvested from the femurs and tibias of 4-to-6-week-old male Sprague-Dawley rats. Cell Counting kit-8 assay was used to determine cytotoxicity of naringenin. Alkaline phosphatase (ALP) activity was measured in cell's precipitates and alizarin-red staining was performed to determine the osteogenic differentiation capacity of the BMSCs. Real-time polymerase chain reaction, enzyme-linked immunosorbent assay and western blotting were adopted to determine the expression of genes and proteins.

Results: The cellular morphology was spindle-shaped, and arranged in radial and whorled patterns. The flow cytometric analysis have confirmed the presence of characteristic surface proteins in the harvested BMSCs. Different concentrations (0-200 μg/ml) of naringenin have no influence on the viability and proliferation rate of the BMSCs. The highest ALP activity was found at culture day 7 and 9 when the concentration of naringenin was 75 and 100 μg/ml. Positive red or dark red stained cells with mineralized nodules can be observed on day 14. The expression of ALP, Runt-related transcription factor 2, CXCR4 and SDF-1a at the gene and protein levels in naringenin-treated cells were significantly higher than those in the control cells. Moreover, AMD3100, an inhibitor of CXCR4, suppressed the expression of the studied genes and proteins.

Conclusions: Naringenin does not show toxic effect on BMSCs. Naringenin promotes the expression of the SDF-1a gene and protein via the SDF-1/CXCR4 signaling pathway. A better understanding of the mechanisms of naringenin action would be helpful for developing specific therapeutic strategies to improve bone regeneration after injuries.

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柚皮苷促进 BMSCs 成骨分化中的 SDF-1/CXCR4 信号通路

引言柚皮苷是一种具有趋化活性的二氢类黄酮化合物，在修复骨组织方面具有良好的应用前景，但其在骨再生，尤其是干细胞成骨分化方面的具体机制有待进一步研究。本研究旨在探讨柚皮素对骨髓间充质干细胞（BMSCs）成骨分化的影响及其在骨髓间充质干细胞C-X-C趋化因子受体4型/基质细胞衍生因子1（SDF-1/CXCR4）信号通路中的作用：从4-6周大的雄性Sprague-Dawley大鼠的股骨和胫骨中采集骨髓间充质干细胞。使用细胞计数试剂盒-8测定柚皮苷的细胞毒性。测定细胞沉淀物中碱性磷酸酶（ALP）的活性，并进行茜素红染色以确定 BMSCs 的成骨分化能力。采用实时聚合酶链反应、酶联免疫吸附试验和免疫印迹法测定基因和蛋白质的表达：结果：细胞形态呈纺锤形，呈放射状和轮状排列。流式细胞分析证实，收获的 BMSCs 中存在特征性表面蛋白。不同浓度（0-200 μg/ml）的柚皮苷对 BMSCs 的活力和增殖率没有影响。当柚皮苷浓度为 75 和 100 μg/ml 时，培养第 7 天和第 9 天的 ALP 活性最高。在第 14 天，可观察到红色或暗红色染色细胞，并伴有矿化结节。柚皮素处理细胞中的 ALP、Runt 相关转录因子 2、CXCR4 和 SDF-1a 在基因和蛋白水平上的表达均明显高于对照组细胞。此外，CXCR4 的抑制剂 AMD3100 也抑制了所研究基因和蛋白质的表达：结论：柚皮素对 BMSCs 没有毒性作用。结论：柚皮素对 BMSCs 没有毒性作用，柚皮素通过 SDF-1/CXCR4 信号通路促进 SDF-1a 基因和蛋白的表达。更好地了解柚皮素的作用机制将有助于开发特定的治疗策略，改善损伤后的骨再生。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Accounts of Chemical Research 化学-化学综合

CiteScore

31.40

自引率

1.10%

发文量

312

审稿时长

2 months

期刊介绍： Accounts of Chemical Research presents short, concise and critical articles offering easy-to-read overviews of basic research and applications in all areas of chemistry and biochemistry. These short reviews focus on research from the author’s own laboratory and are designed to teach the reader about a research project. In addition, Accounts of Chemical Research publishes commentaries that give an informed opinion on a current research problem. Special Issues online are devoted to a single topic of unusual activity and significance. Accounts of Chemical Research replaces the traditional article abstract with an article "Conspectus." These entries synopsize the research affording the reader a closer look at the content and significance of an article. Through this provision of a more detailed description of the article contents, the Conspectus enhances the article's discoverability by search engines and the exposure for the research.