miR-144在人牙周膜成纤维细胞中调控Toll样受体2及骨唾液酸蛋白的表达  

miR-144 regulates TLR2 and BSP expression in human periodontal ligament fibroblasts

作　　者：魏蓉 张源 李新月[2] WEI Rong;ZHANG Yuan;LI Xinyue(Department of Stamotology,School of Medicine,Nankai University,Tianjin 300071,China)

机构地区：[1]南开大学医学院口腔医学系,天津300071 [2]天津市口腔医院牙周病科,天津300041

出　　处：《口腔医学》2020年第7期612-617,共6页Stomatology

摘　　要：目的检测与Toll样受体2(Toll-like receptor 2,TLR2)呈负相关性的微小RNA(microRNA,miRNA)在人牙周膜成纤维细胞(human periodontal ligament fibroblasts,hPDLFs)中与TLR2的作用关系及其对骨唾液酸蛋白(bone sialoprotein,BSP)的调控作用。方法查找与TLR2具有靶向结合关系且呈负相关性的miRNA(miR-23a、miR-144、miR-19a),Real-time PCR方法检测正常及牙龈卟啉单胞菌(Porphyromonas gingivals,P.gingivalis)胞壁成分脂多糖(lipopolysaccharide,LPS)诱导下hPDLFs中目标miRNA的mRNA表达水平;分空白对照组、阴性对照(LPS)组、miRNA过表达对照组、miRNA过表达组、miRNA抑制对照组、miRNA抑制组,用Lipofectamine 2000分别转染hPDLFs,Real-time PCR及Western Blot法分别测定TLR2、BSP的mRNA及蛋白表达水平。结果经LPS诱导后,hPDLFs中TLR2的mRNA表达明显上调(P=0.000),同时miR-144的mRNA下调(P=0.000)。过表达miR-144的hPDLFs在LPS诱导下TLR2的mRNA(P=0.007)和蛋白的表达水平明显下调;而抑制miR-144后TLR2的mRNA(P=0.017)和蛋白表达明显升高。LPS诱导后,hPDLFs中BSP的mRNA(P=0.016)及蛋白表达下调;过表达miR-144的hPDLFs在LPS诱导下BSP的mRNA(P=0.018)和蛋白的表达水平明显上调;而抑制miR-144后BSP的mRNA(P=0.021)和蛋白表达明显下调。结论在hPDLFs中,miR-144具有抑制TLR2和上调BSP表达的作用,与LPS具有负性拮抗作用。Objective To investigate the regulation of miRNA which is negatively correlated with TLR2 on TLR2 and BSP in hPDLFs.Methods miRNAs(miR-23 a,miR-144,miR-19 a)which have targeted bonding relations and have negative correlation with TLR2 were searched.The miRNA mRNA level induced by normal and LPS in hPDLFs was detected by Real-time PCR.The blank control,the negative control,the miRNA mimics,miRNA ASO,as well as their counterpart negative control groups were transfected into cultured hPDLFs by Lipofectamine2000.mRNA and protein expression levels of TLR2 and BSP were detected by Real-time PCR and Western Blot.Results It was shown that TLR2 mRNA expression was up-regulated(P=0.000),while miR-144 mRNA level was down-regulated(P=0.000).Up-regulation of miR-144 could decrease the mRNA(P=0.007)and protein expression levels of TLR2,while the inhibition of miR-144 had the opposite effect(P=0.017).BSP expression was down-regulated after being induced by LPS(P=0.016).Up-regulation of miR-144 could increase the mRNA(P=0.018)and protein expression levels of BSP,while the inhibition of miR-144 played the opposite role(P=0.021).Conclusion MiR-144 can inhibit TLR2 and upregulate BSP expression,and has a reverse effect on LPS,which can counteract the activation of TLR2 and inhibition of BSP by LPS in hPDLFs.

关 键 词：miR-144 TOLL样受体2 骨唾液酸蛋白 牙龈卟啉单胞菌 脂多糖 

分 类 号：R781.4[医药卫生—口腔医学]