miR-181a调控骨髓间充质干细胞中OPG水平及对破骨细胞活性的影响  

miR-181a Reguates the Levels of OPG Protein and Its Effect on the Survival of Osteoclast

作　　者：高敬[1,2,3] 邵秉一[1,2,3] Gao Jing Shao Bingyi(Department of Endodontics, Affiliated Stomatological Hospital, Chongqing Medical University, Chongqing 401147, China Chongqing Key Laboratory of Oral Diseases and Biomedical Sciences, Chongqing 401147, China Chongqing Municipal Key Laboratory of Oral Biomedical Engineering of Higher Education, Chongqing 401147, China)

机构地区：[1]重庆医科大学附属口腔医院牙体牙髓科,重庆401147 [2]口腔疾病与生物医学重庆市重点实验室,重庆401147 [3]重庆市高校市级口腔生物医学工程重点实验室,重庆401147

出　　处：《中国细胞生物学学报》2017年第1期44-51,共8页Chinese Journal of Cell Biology

基　　金：国家自然科学基金面上项目(批准号:31571508;31371473);重庆市医学重点学科建设经费基金<牙体牙髓病学>(2011)资助的课题~~

摘　　要：该研究预测并验证骨髓间充质干细胞(bone marrow mesenchymal stem cells,BMMSCs)中mi R-181a对靶分子骨保护素(osteoprotegerin,OPG)m RNA水平的调控作用,探讨其在骨质疏松发病过程中对破骨细胞活性的影响。通过生物信息学预测mi R-181a可能调控的靶基因OPG,构建含mi R-181a结合位点的OPG m RNA 3′UTR(3′untranslated regions)荧光素酶报告载体。通过双荧光素酶载体系统检测mi R-181a与OPG m RNA 3′UTR相互作用对荧光素酶活性的影响;转染mi R-181a进入BMMSCs并与破骨细胞共培养,TRAP(tartrate resistant acid phosphatase)染色检测共培养体系中破骨细胞数目的改变。结果表明,双荧光素酶报告提示,mi R-181a的靶分子为OPG m RNA,上调mi R-181a(mimics组)水平后,破骨细胞数目较对照组显著增多(P<0.05);下调mi R-181a(inhibitor组)水平后,破骨细胞数目较对照组显著降低(P<0.05)。研究结果显示,mi R-181a可以负调控OPG的蛋白水平,进而影响破骨细胞活性。The research group predict and verify the target molecule osteoprotegerin（OPG） m RNA of mi R-181 a in BMMSCs and investigate the effect of OPG on osteoclast activity in the pathogenesis of osteoporosis. The putative target m RNA of mi R-181 a were predicted by bioinformatics approach. The dual luciferase vector containing 3′ untranslated regions（3′UTR） of OPG m RNA was constructed, and the 3′UTR was regarded as the binding site of mi R-181 a. The interaction between mi R-181 a and target m RNA was verified by dual luciferase activity analysis. mi R-181 a was transfected into BMMSCs and co-cultured with osteoclast, and TRAP staining was used to detect the number of osteoclast in co-culture system. The results of dual luciferase reporter gene assay indicated that mi R-181 a regulated OPG by binding to the 3′UTR of OPG m RNA. In vitro co-culture assays confirmed that BMMSCs transfected with an mi R-181a/mimics increased osteoclast number, whereas BMMSCs/inhibitor reduced osteoclast number. We concluded that mi R-181 a down-regulates the level of OPG protein which might be an important factor to affect the activity of osteoclast.

关 键 词：miR-181a 骨髓间充质干细胞 破骨细胞 

分 类 号：R580[医药卫生—内分泌]