破骨细胞释放凋亡小体微小RNA-30a对成骨活性的影响

doi:10.16098/j.issn.0529-1356.2021.04.010

解剖学报 ›› 2021, Vol. 52 ›› Issue (4): 561-566.doi: 10.16098/j.issn.0529-1356.2021.04.010

破骨细胞释放凋亡小体微小RNA-30a对成骨活性的影响

符义亮¹ 袁凤来^2*

1. 安徽省庐江县人民医院骨科，安徽庐江 231500； 2. 江南大学附属医院中西医结合研究所，江苏无锡 214041

收稿日期:2020-04-01 修回日期:2020-06-12 出版日期:2021-08-06 发布日期:2021-08-06
通讯作者: 袁凤来 E-mail:bjjq88@163.com
基金资助:
江苏省自然科学基金

Effect of osteoclast-derived apoptotic body microR-30a on osteogenic activity

1.Orthopaedics Department, People’s Hospital of Lujiang County, Anhui Lujiang 231500, China; 2.Institute of Integrated Chinese and Western Medicine, the Affiliated Hospital of Jiangnan University, Jiangsu Wuxi 214041, China

Received:2020-04-01 Revised:2020-06-12 Online:2021-08-06 Published:2021-08-06
Contact: YUAN Feng-lai E-mail:bjjq88@163.com

摘要/Abstract

摘要：

目的探讨破骨细胞凋亡释放凋亡小体介导成骨活性的作用。方法通过小鼠（n=10）骨髓单核细胞体外诱导破骨细胞，用抗酒石酸酸性磷酸酶(TRAP)染色和细胞骨架F-actin与DAPI双标免疫荧光鉴定破骨细胞，破骨细胞与小鼠成骨细胞MC-3T3E1共培养体系，DNA片段化ELISA分析破骨细胞凋亡，凋亡小体标志物检测，骨形成标志物Real-time PCR分析，凋亡小体微小RNA (miRNA)表达谱芯片筛查。结果 100 μmol/L 阿伦膦酸钠(ALN) 诱导成熟破骨细胞凋亡，并释放凋亡小体；Western blotting检测表明，ALN诱导凋亡小体特异性表面标志物蛋白表达增强；破骨细胞凋亡小体蛋白C3b表达增高与成骨细胞活性负相关；凋亡小体表达谱芯片筛查提示，miR-30a与骨形成标志物血清碱性磷酸酶(ALP)相关。结论破骨细胞凋亡释放的凋亡小体携带miR-30a抑制成骨细胞活性，凋亡小体可能参与破骨细胞与成骨细胞的对话。

关键词: 破骨细胞, 凋亡小体, 成骨细胞, 骨质疏松症, 免疫印迹法, 小鼠

Abstract:

Objective To explore that whether apoptotic bodies released by osteoclasts mediate osteogenic activity. Methods The osteoclasts were induced from mouse（n=10）bone marrow monocytes in vitro, and were identified by tartrate resistant acid phosphatase (TRAP) staining, F-actin, and DAPI double labeling immunofluorescence. The Co-culture system of osteoclasts and mouse osteoblasts MC-3T3E1 was established. The apoptosis of osteoclasts was analyzed by DNA fragment ELISA. Immunoblotting of apoptotic body markers was investigated. Real-time PCR analysis of bone formation markers was tested. MiRNA expression profiling of apoptotic body was identisfied. Results Alendronate (ALN) 100 μmol/L induced osteoclast apoptosis and caused apoptotic body release from osteoclasts. The expression of C3b and annexin Ⅴ protein was enhanced by ALN; the expression of C3b in osteoclasts was negatively correlated with the activity of osteoblasts; the microarray screening of apoptotic body showed that miR-30a was correlated with bone formation markers and serum alkaline phosphatase (ALP). Conclusion Osteoclast-derived apoptotic body miR-30a can inhibit the activity of osteoblasts. Apoptotic body may participate in the dialogue between osteoclasts and osteoblasts.

Key words: Osteoclast, Apoptotic body, Osteoblast, Osteoporosis, Western blotting, Mouse

中图分类号:

Q291

符义亮袁凤来 . 破骨细胞释放凋亡小体微小RNA-30a对成骨活性的影响[J]. 解剖学报, 2021, 52(4): 561-566.

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破骨细胞释放凋亡小体微小RNA-30a对成骨活性的影响

Effect of osteoclast-derived apoptotic body microR-30a on osteogenic activity

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