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

摘要/Abstract

摘要： 目的检测与TLR2呈负相关性的miRNA在hPDLFs中与TLR2的作用关系及其对骨唾液酸蛋白（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）；（2）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；（3）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具有反向作用，能够抵消LPS对 hPDLFs中TLR2的激活和对BSP的抑制。

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

Abstract: Objective To investigate the regulation of miRNA which is negatively correlated with TLR2 on TLR2 and BSP in hPDLFs.Methods Find miRNAs which target and have negative correlation with TLR2;The miRNA mRNA level induced by normal and LPS in hPDLFs were detected by Real-time PCR;The blank contol,the negtive control ,the miRNA mimics,miRNA ASO,as well as their counterpart negtive control group were transfected into cultured hPDLFs by Lipofectamine2000,the mRNA and protein expression were detected by Reat-time PCR and Western Blot.Results It was shown that TLR2 mRNA expression was up-regulated(P=0.000),while the 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 of TLR2,while the inhibition of miR-144 had the opposite effect(P=0.017).The BSP expression was down-regulated after induced by LPS(P=0.016); Up-regulation of miR-144 could increase the mRNA(P=0.018) and protein expression 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,has a reverse effect with LPS,which can counteract the activation of TLR2 and inhibition of BSP by LPS in hPDLFs.

Key words: miR-144, Toll-like receptor 2, bone sialoprotein, Porphyromonas gingivals, lipopolysaccharide

中图分类号:

R781.4

魏蓉李新月张源. 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[J]. 口腔医学, 2020, 40(7): 612-617.

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