miR-124靶向OSX对牙周膜干细胞成骨分化能力的调控作用  

Study on the regulation of osteogenic differentiation of periodontal ligament stem cells by miR-124 targeting OSX

作　　者：王献刚[1] 冯保静[1] WANG Xian-gang;FENG Bao-jing(Department of Prosthodontic,the Six-People’s Hospital of Anyang City,Henan 455000,China)

机构地区：[1]安阳市第六人民医院,河南455000

出　　处：《中华老年口腔医学杂志》2020年第2期70-75,共6页Chinese Journal of Geriatric Dentistry

摘　　要：目的:探索miR-124对人牙周膜干细胞(periodontal ligament stem cells,PDLSCs)成骨分化能力的影响及调控机制。方法:分离、纯化培养人PDLSCs,检测牙PDLSCs成骨诱导前后miR-124和Osterix(OSX)的表达;转染miR-124沉默(inhibitor)及过表达模拟物(mimics);实时定量PCR(q PCR)检测miR-124和OSXm RNA水平的表达;Western blot检测OSX蛋白水平的变化,茜素红染色分析PDLSCs成骨分化能力的变化;生物信息学手段分析miR-124的靶基因,并通过双荧光素酶报告实验进行验证;共转染miR-124 inhibitor和OSX小干扰RNA(siRNA),检测共转染后对PDLSCs成骨分化能力及OSX表达的影响。结果:成骨分化诱导后PDLSCs中miR-124的表达明显下调,而OSX的表达则显著升高;过表达miR-124可明显抑制OSX的表达和PDLSCs的成骨分化能力,抑制miR-124的表达,则结果相反;生物信息学手段分析OSX为miR-124的潜在靶基因,双荧光素酶报告实验验证miR-124的靶基因为OSX;沉默OSX可逆转miR-124 inhibitor对PDLSCs成骨分化能力的促进作用。结论:miR-124可靶向调控OSX对PDLSCs成骨分化能力产生抑制作用,提示miR-124在PDLSCs组织修复再生过程中发挥重要作用。Objective:To investigate regulation of miR-124 on the osteogenic differentiation of PDLSCs in vitro.Methods:Periodontal ligament stem cells(PDLSCs)were isolated from human healthy periodontal ligament and infected with miR-124 inhibitor and mimics.The expression of miR-124 and OSX were detected in PDLSCs after osteogenic differentiation.Then the osteogenic differentiation was detected by alizarin red assay.The expression of miR-124 and OSX were evaluated by q-RCR and Western blot.The potential target gene of miR-124 was verified by dual luciferase report.Results:The expression of OSX were significant up-regulated but miR-124 were down-regulated after osteogenic differentiation.Overexpression of mir-124 significantly inhibited the expression of OSX and osteogenic differentiation of PDLSCs,and the opposite was observed if the expression of mir-124 was inhibited OSX was the potential target gene of miR-124,which analyzed by bioinformatics and verified dual luciferase report.In addition,si-OSX was able to reverse the promoted effect of miR-124 inhibitor on the osteogenic differentiation of PDLSCs.Conclusion:miR-124 inhibit the osteogenic differentiation ability of PDLSCs via inhibiting the expression of OSX,which was implied that miR-124 plays an important role in PDLSCs involved in injury repair.

关 键 词：miR-124 牙周膜干细胞(PDLSCs) 成骨分化 OSX 

分 类 号：R781.4[医药卫生—口腔医学]