牵张刺激下牙周膜细胞外泌体miR-181d-5p在调控成骨细胞分化中的作用  

The Role of Exosome MiR-181d-5p Secreted by Periodontal Ligament Cells under Stretch Loading in the Regulation of Osteoblast Differentiation

作　　者：张轶凡 瞿方 王莹莹[1] 吴雅琴[1] 宋迎爽 曹希萌 沈荧怡 胥春 ZHANG Yifan;QU Fang;WANG Yingying;WU Yaqin;SONG Yingshuang;CAO Ximeng;SHEN Yingyi;XU Chun(Department of Prosthodontics,Shanghai Ninth People’s Hospital,Shanghai Jiao Tong University School of Medicine/College of Stomatology,Shanghai Jiao Tong University/National Center for Stomatology/National Clinical Research Center for Oral Diseases/Shanghai Key Laboratory of Stomatology/Shanghai Research Institute of Stomatology,Shanghai 200011,China)

机构地区：[1]上海交通大学医学院附属第九人民医院口腔修复科、上海交通大学口腔医学院、国家口腔医学中心、国家口腔疾病临床医学研究中心、上海市口腔医学重点实验室、上海市口腔医学研究所,上海200011

出　　处：《医用生物力学》2023年第5期910-916,共7页Journal of Medical Biomechanics

基　　金：国家自然科学基金项目(82071157,31470903,31270991,30900282);上海市卫生健康委员会卫生行业临床研究专项资助项目(201940009);上海交通大学医学院附属第九人民医院原创探索项目(JYYC002)。

摘　　要：目的探索牵张加载下牙周膜细胞(periodontal ligament cells,PDLCs)外泌体对成骨细胞分化的影响,以及牵张加载下PDLCs外泌体在力相关牙周组织改建中的调控作用。方法对体外培养人PDLCs施加20%牵张应变后,提取上清中的外泌体与成骨细胞共培养,检测成骨细胞分化情况,并探索外泌体中发挥调控作用的miRNA。结果牵张加载下PDLCs分泌的外泌体可上调成骨细胞中碱性磷酸酶(alkaline phosphatase,ALP)、转录因子Osterix(Osx)、I型胶原蛋白(type I collagen,COL-1)、Runt相关转录因子2(Runt-related transcription factor 2,Runx2)、骨钙素(osteocalcin,OCN)和骨形态发生蛋白2(bone morphogenetic protein 2,BMP 2)等成骨相关因子表达,促进细胞成骨向分化。牵张加载下PDLCs外泌体差异表达miRNA中,miRNA-181d-5p表达量上调达107倍,将其转染成骨细胞后,成骨细胞中ALP、Osx、Col-1、Runx2、Ocn和BMP2等成骨相关因子表达亦增加。结论本研究初步揭示了牵张加载下PDLCs外泌体miR-181d-5p在调控成骨细胞分化中的作用,对于明确力相关牙周组织改建中的信号通路具有重要科学意义。Objective To investigate the effect of exosomes,derived from periodontal ligament cells(PDLCs)under stretch loading on osteoblast differentiation,and the role of these exosomes in force-related periodontal 910 tissue remodeling.Methods After 20%strain was applied to human PDLCs cultured in vitro,exosomes from the supernatant were extracted and co-cultured with osteoblasts,then the differentiation of osteoblasts was detected,and miRNA which played a regulatory role in exosomes was explored.Results Exosomes secreted by PDLCs under stretch loading could up-regulate the expression of osteogenic-related factors including alkaline phosphatase(ALP),transcription factor osterix(osterix,Osx),type I collagen(Col-1),Runt-related transcription factor 2(Runx2),osteocalcin(Ocn),and bone morphogenetic protein 2(BMP2)in osteoblasts,as well as promote osteogenic differentiation of cells.Among the miRNAs differentially expressed in PDLCs exosomes under stretch loading,the expression of miRNA-181d-5p was up-regulated by 107 times,and the expression of osteogenic-related factors including ALP,Osx,Col-1,Runx2,Ocn and BMP2 was also increased in osteoblasts transfected with miRNA-181d-5p.Conclusions This study preliminarily reveals the role of exosome miR-181d-5p secreted by PDLCs under stretch loading in regulating the differentiation of osteoblast,which is of scientific significance for clarifying the signaling pathway in force-related periodontal tissue remodeling.

关 键 词：牙周膜细胞 牵张 外泌体 miR-181d-5p 成骨分化 

分 类 号：R318.01[医药卫生—生物医学工程]