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脂联素对小鼠骨髓来源单核细胞破骨细胞生成的影响。

Effects of adiponectin on osteoclastogenesis from mouse bone marrow-derived monocytes.

作者信息

Wu Xia, Huang Leitao, Liu Jichun

机构信息

Department of Thoracic and Cardiovascular Surgery, The First Affiliated Hospital of Nanchang University, Nanchang, Jiangxi 330006, P.R. China.

Department of Orthopedics, The First Affiliated Hospital of Nanchang University, Nanchang, Jiangxi 330006, P.R. China.

出版信息

Exp Ther Med. 2019 Feb;17(2):1228-1233. doi: 10.3892/etm.2018.7069. Epub 2018 Dec 7.

Abstract

The aim of the present study was to investigate the effects of adiponectin on bone marrow-derived monocytes (BMMs) in the process of osteoclastogenesis. Primary BMMs derived from the mouse bone marrow were cultured, which were then treated with different concentrations of adiponectin and macrophage colony stimulating factor (M-CSF). Cell viability was determined by measuring the absorbance after 24 h with Cell Counting Kit-8 reagent. BMM cells treated with adiponectin and receptor activator of nuclear factor-κB ligand (RANKL) were induced and differentiated to mature osteoclasts for 1 week, and then stained with tartrate-resistant acid phosphatase (TRAP). The number of osteoclasts was evaluated under light microscopy. The expression of adiponectin in BMMs at the gene and protein levels was further assessed with reverse transcription-quantitative polymerase chain reaction and western blotting, respectively. The cellular proliferation experiment demonstrated that the optical density value decreased gradually with an increase of adiponectin concentration, with statistically significant differences detected among groups. In addition, the number of osteoclasts in the adiponectin-treated group was significantly reduced compared with that in the control group. Adiponectin expression was confirmed in BMMs at both the protein and mRNA levels. In conclusion, the present data demonstrated that adiponectin has a significant inhibitory effect on the osteoclast differentiation and proliferation of BMMs, suggesting a novel strategy for preventing osteoporosis.

摘要

本研究的目的是探讨脂联素在破骨细胞形成过程中对骨髓来源单核细胞(BMMs)的影响。培养从小鼠骨髓中分离得到的原代BMMs,然后用不同浓度的脂联素和巨噬细胞集落刺激因子(M-CSF)进行处理。使用细胞计数试剂盒-8试剂在24小时后通过测量吸光度来测定细胞活力。用脂联素和核因子κB受体激活剂配体(RANKL)处理BMM细胞并诱导分化1周,使其成为成熟破骨细胞,然后用抗酒石酸酸性磷酸酶(TRAP)进行染色。在光学显微镜下评估破骨细胞的数量。分别用逆转录定量聚合酶链反应和蛋白质印迹法进一步评估BMMs中脂联素在基因和蛋白质水平的表达。细胞增殖实验表明,随着脂联素浓度的增加,光密度值逐渐降低,各组间差异有统计学意义。此外,与对照组相比,脂联素处理组的破骨细胞数量显著减少。在BMMs中证实了脂联素在蛋白质和mRNA水平均有表达。总之,目前的数据表明脂联素对BMMs的破骨细胞分化和增殖具有显著的抑制作用,提示了一种预防骨质疏松症的新策略。

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