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作 者:钱丽萍 马灵芝 余兵[1] 周治[2] 刘亚丽[1] QIAN Li-ping;MA Ling-zhi;YU Bing;ZHOU Zhi;LIU Ya-li(School of Stomatology, Kunming Medical University,Kunming Yunnan 650500;Department of Orthodontics,The Second People's Hospital of Yunnan Province,Kunming Yunnan 650021,China)
机构地区:[1]昆明医科大学口腔医学院,云南昆明650500 [2]云南省第二人民医院口腔正畸科,云南昆明650021
出 处:《昆明医科大学学报》2018年第12期5-10,共6页Journal of Kunming Medical University
基 金:国家自然科学基金资助项目(31301067);高等学校博士学科点专项基金资助项目(20135317120005);云南省研究基金及科技人才计划基金资助项目(2013Z106;2015HB044;2017FE468-227;D-201667);昆明医科大学研究生创新基金资助项目(2018S171)
摘 要:目的探讨体外炎症微环境中,miR-17对牙周膜干细胞(periodontal ligament stem cells,PDLSCs骨向分化能力的影响。方法体外有限稀释法克隆化和密度梯度离心法分别培养牙周膜干细胞(PDLSCs)和外周血免疫细胞(PBMCs),并建立PDLSCs与免疫细胞的Transwell共培养炎症微环境体系。使用定量PCR技术检测miR-17的表达,并将PDLSCs转染miR-17模拟物Mimics和抑制剂Inhibitor后进行成骨诱导,ALP染色,定量PCR和Western Blot检测细胞成骨分化的能力。结果共培养组PDLSCs中miR-17表达降低,转染Mimics后共培养细胞成骨分化能力升高,转染Inhibitor后细胞成骨分化能力降低。结论炎症微环境中miR-17可以促进PDLSCs骨向分化。Objective To investigate the effects of miR-17 on the osteogenic differentiation potential of human periodontal ligamentcells(PDLSCs)in chronic inflammation microenvironment.Methods PDLSCs and Peripheral Blood Mononuclear Cells(PBMCs)were isolated respectively by the limiting dilution technique and by density gradient centrifugation from healthy individuals.Then a co-culture system of PDLSCs-PBMCs(in vitro)was established.The miR-17 expression was analyzed by real-time RT-PCR.Alkaline phosphatase staining,real-time RT-PCR and Western blot were used to evaluate the osteogenic differentiation potential of PDLSCs after transient transfection of miR-17 mimics or inhibitor.Results The miR-17 expression in the co-culture system of PDLSCs-PBMCs was reduced comparing to the control group,and the osteogenic differentiation potential of PDLSCs-PBMCs was promoted by miR-17 mimics,and reduced by miR-17 inhibitor.Conclusion miR-17 can promote osteogenic differentiation in the co-culture system of PDLSCs-PBMCs.
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