AIM: To study the expression of miR -146a in inflammatory periodontal ligment ctem cells (IPLSCs) and to verify the relationship of miR-146a and TLR4 in the inflammatory mechanism of periodontitis. METHODS:The expression level of miR-146a in PDLSCs and LPS stimulated PDLSCs(inflammatory PDLSCs, IP-DLSCs) was detected by Real-Time PCR.The effects of miR-146a on TLR4 gene expression was verified by Real-Time PCR,Western blot and the dual-luciferase reporter assay. RESULTS:The expression of miR-146a in IPDLSCs was higher than that in PDLSCs(P<0.05),and miR-146a expression was significantly up-regulated by LPS in IP-DLSCs. Up-regulation of miR-146a could suppress TLR4 mRNA and protein expression(P<0.05); inhibition of miR-146a expression could promote TLR4 mRNA and protein expression(P<0.05),the dual-luciferase reporter as-say demonstrated that miR-146a targeted inbibition of the specific 3'untranslated region(3' UTR) sequence of TLR4 gene. CONCLUSION: miR-146a is upregulated in IPDLSCs, and it can directly inhibit TLR4 gene expression. miR-146a may participate in the regulation of inflammatory reaction of IPDLSCs.%目的:检测miR-146a在人健康及脂多糖(LPS)刺激炎症牙周膜干细胞(PDLSCs)后的表达情况,并探讨其与TLR4的作用关系.方法:分别提取牙周健康者和牙周炎患者的PDLSCs,通过Real-Time PCR检测人健康及LPS刺激后炎症PDLSCs中miR-146a 的mRNA表达水平;Real-Time PCR、Western blot及荧光素酶报告基因检测miR-146a与TLR4的相互作用关系.结果:炎症PDLSCs中miR-146a的mRNA表达水平明显高于健康组(P <0. 05);LPS 刺激的炎症 PDLSCs 中 miR-146a 的 mRNA 表达水平显著增高(P<0.05).与空白组相比,过表达miR-146a后,TLR4 mRNA及蛋白的表达水平均明显降低(P<0.05);抑制miR-146a的表达后,TLR4 mRNA及蛋白的表达水平均显著增加(P<0.05);荧光素酶报告基因检测结果显示, miR-146a与TLR4的3′UTR具有靶向抑制作用.结论:miR-146a在人炎症PDLSCs中表达上调,同时可显著下调其靶基因TLR4的表达,推测miR-146a可能参与了DPLSCs的炎症反应调控.
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