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 microRNA-30a on osteogenic activity

作　　者：符义亮 袁凤来 FU Yi-liang;YUAN Feng-lai(Orthopaedics Department,People’s Hospital of Lujiang County,Anhui Lujiang 231500,China;Institute of Integrated Chinese and Western Medicine,the Affiliated Hospital of Jiangnan University,Jiangsu Wuxi 214041,China)

机构地区：[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

出　　处：《解剖学报》2021年第4期561-566,共6页Acta Anatomica Sinica

基　　金：国家自然科学基金(81770876);江苏省自然科学基金(BK20191141)。

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100μmol/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。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 Coculture 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)100μmol/L induced osteoclast apoptosis and caused apoptotic body release from osteoclasts.The expression of C3b and annexinⅤprotein 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.

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 

分 类 号：Q291[生物学—细胞生物学]