摘要： 目的 探讨辛伐他汀抑制破骨细胞分化的机制。方法 RAW264.7细胞培养并传代，sRANKL+M-CSF诱导RAW264.7细胞向破骨细胞分化；WST-1检测不同浓度辛伐他汀对破骨细胞增殖活性的影响；TRAP染色检测辛伐他汀对破骨细胞分化的影响；免疫荧光检测辛伐他汀对破骨细胞肌动蛋白环形成的影响及对通路蛋白p65细胞核内转移的影响；Western blotting检测加入辛伐他汀后对通路蛋白p65的影响。结果 WST-1结果显示，辛伐他汀（10^-6 mol/L）作用24 h后明显抑制了破骨细胞的增殖活性。TRAP染色显示，成熟的破骨细胞增多核，胞浆嗜酸性，染色呈酒红色，加入辛伐他汀及核因子κB（NF-κB）通路抑制PDTC后，破骨细胞的形成及分化明显受到抑制。免疫荧光检测显示，辛伐他汀抑制破骨细胞肌动蛋白环的形成、抑制NF-κB信号通路核心蛋白p65的细胞核内转位。免疫印迹结果显示，辛伐他汀能够抑制p65的磷酸化。结论 辛伐他汀通过NF-κB信号通路抑制破骨细胞的分化。

辛伐他汀通过NF-κB通路抑制破骨细胞的分化

于冬冬¹, 赵丹阳², 杨鸫祥¹, 杨关林³

1. 辽宁中医药大学附属医院骨科, 沈阳 110032;
2. 沈阳市第一人民医院神经内科, 沈阳 110041;
3. 辽宁中医药大学附属医院中西医结合内科, 沈阳 110032

收稿日期:2018-09-03出版日期:2020-03-30发布日期:2020-03-19
通讯作者:杨关林E-mail:guanlinyang@yeah.net
作者简介:于冬冬(1983-),男,副主任医师,博士.
基金资助:国家自然科学基金（81973719）；辽宁省自然科学基金（20170540597）；辽宁省教育厅科学技术研究项目（L201921）；辽宁中医药大学中医脏象理论及应用教育部重点实验室开放基金（zyzx1908）


关键词: 辛伐他汀, 绝经后骨质疏松症, 破骨细胞分化
Abstract: Objective To explore the mechanism through which simvastatin inhibits osteoclast differentiation. Methods RAW264.7 cells were cultured and passaged,and sRANKL+M-CSF was added to induce RAW264.7 cell differentiation into osteoclasts. WST-1 assay was used to detect the effects of simvastatin on the proliferation of osteoclasts exposed to different concentrations of sRANKL+M-CSF. The effect of simvastatin on osteoclast differentiation was detected by TRAP staining. The effects of simvastatin on F-actin ring formation in osteoclasts and nuclear translocation of the nuclear factor kappa B (NF-κB) pathway protein p65 were assessed by immunofluorescence. The effect of simvastatin on p65 protein expression was assessed by western blotting. Results WST-1 assay showed that simvastatin (10^-6 mol/L) significantly inhibited osteoclast proliferation after 24 h. TRAP staining showed that mature osteoclasts were nucleocytic,with eosinophilic cytoplasm that stained a wine red color. Adding simvastatin together with PDTC (an inhibitor of the NF-κB pathway) noticeably inhibited formation and differentiation of osteoclasts. Immunofluorescence assays showed that simvastatin inhibited the formation of actin rings in osteoclasts,and inhibited nuclear translocation of the core protein p65 of the NF-κB signaling pathway. Western blotting revealed that simvastatin could inhibit p65 phosphorylation. Conclusion Simvastatin inhibits osteoclast differentiation through the NF-κB pathway.
Key words: simvastatin, postmenopasal osteoporosis, osteoclast differentiation
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