miR-381-3p在炎性环境下对人牙周膜干细胞成骨分化的调节作用及机制  

The regulatory effect and mechanism of miR-381-3p on osteogenic differentiation of human periodontal membrane stem cells in inflammatory environment

作　　者：黄亿亿 汪青风 戴卓 HUANG Yiyi;WANG Qingfeng;DAI Zhuo(The Affiliated Stomatological Hospital of Nanjing University,Nanjing Stomatological Hospital,Nanjing 210000,China)

机构地区：[1]南京大学附属口腔医院·南京市口腔医院,江苏南京210000

出　　处：《西部医学》2024年第9期1282-1287,1294,共7页Medical Journal of West China

基　　金：南京市卫生科技发展专项资金项目(YKK22182)。

摘　　要：目的探讨miR-381-3p对炎性环境下人牙周膜干细胞(hPDLSCs)成骨分化的促进作用及机制。方法体外分离和培养hPDLSCs细胞,用流式细胞法对hPDLSCs细胞进行鉴定。对hPDLSCs细胞进行miR-381-3p转染并用双萤光素酶检测试剂分析miR-381-3p的目的基因。将hPDLSCs细胞分为对照组、miR-NC组、miR-NC+LPS组、miR-381-3p抑制物组、miR-381-3p抑制物+LPS组、miR-381-3p模拟物组和miR-381-3p模拟物+LPS组。干预后检测hPDLSCs成骨分化能力及hPDLSCs细胞中ALP、miR-381-3p、TLR4、NF-κB和Runx2 mRNA的表达水平。结果分离的细胞符合间充质干细胞特征;miR-381-3p对TLR4有潜在的结合位点,存在靶向调节作用;与对照组比较,LPS组hPDLSCs细胞中miR-381-3p、TLR4和NF-κB mRNA表达增加,ALP、矿化结节形成量和Runx2 mRNA表达降低(P<0.05);与miR-NC组和miR-NC+LPS组比较,miR-381-3p抑制物组和miR-381-3p抑制物+LPS组hPDLSCs细胞中miR-381-3p、TLR4和NF-κB mRNA表达降低,ALP、矿化结节形成量和Runx2 mRNA表达增加,miR-381-3p抑制物组变化更显著(P<0.05);与其他组比较,miR-381-3p模拟物组和miR-381-3p模拟物+LPS组hPDLSCs细胞中miR-381-3p、TLR4和NF-κB mRNA表达增加,ALP、矿化结节形成量和Runx2 mRNA表达降低,miR-381-3p模拟物+LPS组变化更显著(P<0.05)。结论在炎性环境下,hPDLSCs细胞中miR-381-3p表达增加。转染miR-381-3p抑制物可以抑制TLR4/NF-κB信号通路的激活,促进hPDLSCs细胞成骨分化。Objective To explore the promoting effect and mechanism of miR-381-3p on osteogenic differentiation of human periodontal membrane stem cells(hPDLSCs)in inflammatory environment.Methods hPDLSCs were isolated and cultured in vitro,and identified by flow cytometry.hPDLSCs cells were transfected with miR-381-3p and the target gene of miR-381-3p was analyzed with bifluciferase detection reagent.hPDLSCs cells were divided into control group,miR-NC group,miR-NC+LPS group,miR-381-3p inhibitor group,miR-381-3p inhibitor+LPS group,miR-381-3p simulator group and miR-381-3p simulator+LPS group.After the intervention,the osteogenic differentiation ability of hPDLSCs and the ALP,mRNA expression of miR-381-3p,TLR4,NF-κB and Runx2 in hPDLSCs were detected.Results The isolated cells fit the characteristics of mesenchymal stem cells.miR-381-3p had a potential binding site for TLR4 and had a targeted regulatory effect.Compared with the control group,the mRNA expressions of miR-381-3p,TLR4 and NF-κB in hPDLSCs cells in LPS group were increased,while the levels of ALP,mineralized nodule formation and Runx2 mRNA were decreased(P<0.05).Compared with miR-NC group and miR-NC+LPS group,mRNA expressions of miR-381-3p,TLR4 and NF-κB in hPDLSCs cells of miR-381-3p inhibitor group and miR-381-3p inhibitor+LPS group decreased,while the levels of ALP,mineralized nodule formation and Runx2 mRNA were increased,and the changes of miR-381-3p inhibitor group were more significant(P<0.05).Compared with other groups,the mRNA expressions of miR-381-3p,TLR4 and NF-κB in hPDLSCs cells of miR-381-3p mimics group and miR-381-3p mimics+LPS group were increased,while the levels of ALP,mineralized nodule formation and Runx2 mRNA were decreased,and the changes of miR-381-3p mimics+LPS group were more significant(P<0.05).Conclusion In inflammatory environment,the expression of miR-381-3p increases in hPDLSCs.Transfection of miR-381-3p inhibitor can inhibit the activation of TLR4/NF-κB signaling pathway and promote the osteogenic differentiation of hPDLSCs.

关 键 词：miR-381-3p 炎症 人牙周膜干细胞 成骨分化 TLR4/NF-κB信号通路 

分 类 号：R781.4[医药卫生—口腔医学]