MLO-Y4来源外泌体对破骨细胞调控机制的研究  

Mechanism of re gulation of oste oclasts by MLO-Y4-derived exosomes

作　　者：赵丽洲 勾蓉 王晨 涂小林[1] ZHAO Lizhou;GOU Rong;WANG Chen;TU Xiaolin(Laboratory of Bone Development and Regeneration,Institute of Life Sciences,Chongqing Medical University,Chongqing 400016,China;Health Examination Center,the Affiliated Cancer Hospital of Chongqing University,Chongqing 400030,China;Southwest Hospital,Army Medical University,Chongqing 400038,China)

机构地区：[1]重庆医科大学生命科学研究院骨发育与再生实验室,重庆400016 [2]重庆大学附属肿瘤医院体检中心,重庆400030 [3]陆军军医大学西南医院,重庆400038

出　　处：《中国骨质疏松杂志》2023年第9期1255-1260,共6页Chinese Journal of Osteoporosis

基　　金：国家自然科学基金(82072450)。

摘　　要：目的探讨骨细胞系MLO-Y4来源的外泌体对破骨细胞的调控机制。方法收集MLO-Y4细胞的培养基,通过超高速离心法获得MLO-Y4细胞外泌体(MLO-Y4-Exo),Western blot和透射电镜鉴定其特征;MLO-Y4-Exo与骨髓巨噬细胞共培养,TRAP染色检测其对破骨细胞分化的作用;小鼠顶骨皮下注射MLO-Y4-Exo检测其对体内破骨细胞分化的影响;通过小干扰RNA检测MLO-Y4-Exo对促破骨分化的机制。结果MLO-Y4外泌体大小形态满足外泌体特征,高表达CD63和Alix外泌体蛋白;MLO-Y4-Exo在体外和体内均促进破骨细胞分化(P<0.05);RANKL小干扰RNA实验证实MLO-Y4-Exo通过向破骨前体细胞传递RANKL蛋白促进破骨细胞分化(P<0.05)。结论骨细胞系MLO-Y4来源的外泌体通过向破骨前体细胞传递促破骨分化因子RANKL促进破骨细胞生成。Objective To investigate the effect of osteocyte-like cell MLO-Y4-derived exosomes on osteoclast differentiation.Me thods Osteocyte-like cell MLO-Y4-derived exosomes(MLO-Y4-Exo)were obtained by ultrahigh speed centrifugation.Their characteristics were identified using Western blotting and transmission electron microscopy.MLO-Y4-Exo were co-cultured with bone marrow macrophages.TRAP staining was conducted to detect the osteoclast differentiation.Mice were subjected to subcutaneous supra-calvarial injection of MLO-Y4-Exo for 5 consecutive days to detect the osteoclast differentiation in vivo.The mechanism of MLO-Y4-Exo promoting osteoclast differentiation was detected with small interfering RNA.Re sults The size and morphology of MLO-Y4-Exo met the exosome characteristics and highly expressed CD63 and Alix exosome proteins.MLO-Y4-Exo promoted osteoclast differentiation in vitro and in vivo(P<0.05).RANKL small interfering RNA assay confirmed that MLO-Y4-Exo promoted osteoclast differentiation by transmitting RANKL protein to osteoclast precursors(P<0.05).Conclusion MLO-Y4-derived exosomes promote osteoclast formation by transferring RANKL to osteoclast precursors.

关 键 词：MLOY-4 外泌体 破骨细胞 RANKL 

分 类 号：R336[医药卫生—人体生理学] R392-33[医药卫生—基础医学]