上调miR-31表达通过Wnt/β-catenin信号通路对牙髓干细胞成骨分化的影响  

Effect of up-regulation of miR-31 expression on osteogenic differentiation of dental pulp stem cells through Wnt-β/catenin signaling pathway

作　　者：张雅琪 米靖 杨景荣 李欣明 李利 ZHANG Yaqi;MI Jing;YANG Jingrong;LI Xinming;LI Li(Department of Endodontics,Tianjin Stomatological Hospital,School of Medicine,Nankai University,Tianjin 300041,China;Tianjin Key Laboratory of Oral and Maxillofacial Function Reconstruction,Tianiin 300041,China;Department of Stomatology,Qingxian People’s Hospital,Cangzhou 062650,China)

机构地区：[1]南开大学医学院天津市口腔医院牙体牙髓病一科,天津300041 [2]天津市口腔功能重建重点实验室,天津300041 [3]河北省青县人民医院口腔科,河北沧州062650

出　　处：《吉林大学学报(医学版)》2025年第2期412-419,共8页Journal of Jilin University:Medicine Edition

基　　金：天津市卫健委科技项目(TJWJ2024QN073)。

摘　　要：目的:探讨微小RNA(miRNA)-31对牙髓干细胞(DPSCs)成骨分化的影响,并阐明其可能的作用机制。方法:将对数生长数期DPSCs分为对照组(不处理)、NC组(转染随机序列对照物miRNA)、Agomir组(转染miR-31模拟物agomiR-31)和联合组(转染miR-31模拟物agomiR-31m同时加入XAV939)。转染48 h后采用实时荧光定量PCR(RT-qPCR)法检测各组DPSCs中miR-31表达水平,MTT法检测各组DPSCs增殖能力,茜素红染色法检测各组DPSCs中钙质沉积,碱性磷酸酶(ALP)染色法检测各组DPSCs成骨分化程度,Western blotting法检测各组DPSCs无翅型MMTV整合位点家族蛋白(Wnt)/β-连环蛋白(β-catenin)信号通路相关蛋白表达水平。结果:对照组和NC组DPSCs中miR-31表达水平,24、48和72 h时细胞增殖能力,钙化区面积构成比和ALP活性比较,差异均无统计学意义(P>0.05)。与对照组和NC组比较,Agomir组DPSCs中miR-31表达水平,24、48和72h时细胞增殖能力,钙化区面积构成比和ALP活性均明显升高(P<0.05)。与Agomir组比较,联合组DPSCs中miR-31表达水平,24、48和72h时细胞增殖能力,钙化区面积构成比和ALP活性均明显降低(P<0.05)。对照组和NC组DPSCs中糖原合成酶激酶3β(GSK-3β)、β-catenin和Runt相关转录因子2(Runx2)蛋白表达水平比较差异均无统计学意义(P>0.05);与对照组和NC组比较,Agomir组DPSCs中GSK-3β蛋白表达水平降低(P<0.05),β-catenin和Runx2蛋白表达水平升高(P<0.05);与Agomir组比较,联合组DPSCs中GSK-3β蛋白表达水平升高(P<0.05),β-catenin和Runx2蛋白表达水平降低(P<0.05)。结论:上调miR-31可促进DPSCs增殖和成骨分化,其作用机制可能与激活Wnt/β-catenin信号通路有关。Objective:To investigate the effect of up-regulation of microRNA-31(miRNA-31)on the osteogenic differentiation of dental pulp stem cells(DPSCs),and to elucidate its possible mechanism.Methods:The DPSCs in logarithmic growth phase were divided into control group(no treatment),NC group(transfected with random sequence control),Agomir group(transfected with miR-31 mimic agomiR-31),and combination group(transfected with miR-31 mimic agomiR-31 and added with XAV939).After 48 h of transfection,real-time fluorescence quantitative PCR(RT-qPCR)was used to detect the expression levels of miR-31 in the DPSCs in various groups.MTT assay was used to detect the proliferation abilities of the DPSCs in various groups.Alizarin red staining was used to detect calcium deposition in the DPSCs in various groups.Alkaline phosphatase(ALP)staining was used to detect the degree of osteogenic differentiation of the DPSCs in various groups.Western blotting method was used to detect the expression levels of proteins related to the wingless-type MMTV integration site family(Wnt)/β-catenin signaling pathway in the DPSCs in various groups.Results:There were no significant differences in the miR-31 expression level,the cell proliferation abilities at 24,48 and 72 h,the ratio of calcified region,and the ALP ability between control group and NC group(P>0.05).Compared with control group and NC group,the expression level of miR-31,the cell proliferation abilities at 24,48 and 72 h,the ratio of calcified region,and the ALP activity in the DPSCs in Agomir group were increased(P<0.05).Compared with Agomir group,the expression level of miR-31,the cell proliferation abilities at 24,48 and 72 h,the ratio of calcified region,and the ALP activity in the DPSCs in combination group were decreased(P<0.05).There were no significant difference in the expression levels of glycogen synthase kinase 3β(GSK-3β),β-catenin and Runt-associated transcription factor 2(Runx2)in the DPSCs between control group and NC group(P>0.05).Compared with control group and NC gro

关 键 词：微小RNA-31 牙髓干细胞 成骨分化 细胞增殖 Β-连环蛋白 

分 类 号：R781.3[医药卫生—口腔医学]