Abstract：
BACKGROUND: MicroRNAs are differentially expressed in periodontitis tissues. They are involved in cellular responses to inflammation and can be used as markers for diagnosing periodontitis. Microarray analysis showed that the expression level of microRNA-671-5p in periodontal tissues of patients with periodontitis was increased. In this study, we investigated the mechanism of action of microRNA-671-5p in human periodontal ligament stem cells (hPDLSCs) under inflammatory conditions.
RESULTS: HPDLSCs were treated with lipopolysaccharide (LPS) to establish an inflammation model. The cell survival rate was determined using the cell counting kit-8 (CCK8). Real-time quantitative reverse transcription polymerase chain reaction (qRT-PCR) and western blot analyses were used to detect the expression of microRNA-671-5p and dual-specificity phosphatase (DUSP) 8 proteins, respectively, Interleukin (IL)-6, IL-1β, and tumor necrosis factor (TNF)-α were detected using qRT-PCR and Enzyme-linked immunosorbent assay (ELISA). A dual-luciferase reporter system was employed to determine the relationship between micoRNA-671-5p and DUSP8 expression. Activation of the p38 mitogen-activated protein kinase (MAPK) signaling pathway was confirmed using western blot analysis. Following the treatment of hPDLSCs with LPS, the expression levels of microRNA-671-5p in hPDLSCs were increased, cell viability decreased, and the expression of inflammatory factors displayed an increasing trend. MicroRNA-671-5p targets and binds to DUSP8. Silencing microRNA-671-5p or overexpressing DUSP8 can improve cell survival rate and reduce inflammatory responses. When DUSP8 was overexpressed, the expression of p-p38 was reduced.
CONCLUSIONS: microRNA-671-5p targets DUSP8/p38 MAPK pathway to regulate LPS-induced proliferation and inflammation in hPDLSCs.
RESULTS: HPDLSCs were treated with lipopolysaccharide (LPS) to establish an inflammation model. The cell survival rate was determined using the cell counting kit-8 (CCK8). Real-time quantitative reverse transcription polymerase chain reaction (qRT-PCR) and western blot analyses were used to detect the expression of microRNA-671-5p and dual-specificity phosphatase (DUSP) 8 proteins, respectively, Interleukin (IL)-6, IL-1β, and tumor necrosis factor (TNF)-α were detected using qRT-PCR and Enzyme-linked immunosorbent assay (ELISA). A dual-luciferase reporter system was employed to determine the relationship between micoRNA-671-5p and DUSP8 expression. Activation of the p38 mitogen-activated protein kinase (MAPK) signaling pathway was confirmed using western blot analysis. Following the treatment of hPDLSCs with LPS, the expression levels of microRNA-671-5p in hPDLSCs were increased, cell viability decreased, and the expression of inflammatory factors displayed an increasing trend. MicroRNA-671-5p targets and binds to DUSP8. Silencing microRNA-671-5p or overexpressing DUSP8 can improve cell survival rate and reduce inflammatory responses. When DUSP8 was overexpressed, the expression of p-p38 was reduced.
CONCLUSIONS: microRNA-671-5p targets DUSP8/p38 MAPK pathway to regulate LPS-induced proliferation and inflammation in hPDLSCs.
摘要：
背景：MicroRNAs在牙周炎组织中差异表达。它们参与细胞对炎症的反应，可用作诊断牙周炎的标记。基因芯片分析表明，牙周炎患者牙周组织中microRNA-671-5p的表达水平升高。在这项研究中,我们研究了炎症条件下microRNA-671-5p在人牙周膜干细胞（hPDLSCs）中的作用机制。
结果：用脂多糖（LPS）处理HPDLSCs以建立炎症模型。使用细胞计数试剂盒-8(CCK8)测定细胞存活率。实时定量逆转录聚合酶链反应(qRT-PCR)和蛋白质印迹分析用于检测microRNA-671-5p和双特异性磷酸酶(DUSP)8蛋白的表达,分别,白细胞介素（IL）-6，IL-1β，采用qRT-PCR和酶联免疫吸附试验(ELISA)检测肿瘤坏死因子(TNF)-α。使用双荧光素酶报告系统来确定micoRNA-671-5p与DUSP8表达之间的关系。使用蛋白质印迹分析证实p38丝裂原活化蛋白激酶(MAPK)信号通路的激活。用LPS处理hPDLSCs后，microRNA-671-5p在hPDLSCs中的表达水平升高,细胞活力下降，炎症因子的表达呈上升趋势。MicroRNA-671-5p靶向并结合DUSP8。沉默microRNA-671-5p或过表达DUSP8可以提高细胞存活率并减少炎症反应。当DUSP8过表达时，p-p38的表达降低。
结论：microRNA-671-5p靶向DUSP8/p38MAPK通路以调节LPS诱导的hPDLSCs增殖和炎症。
结果：用脂多糖（LPS）处理HPDLSCs以建立炎症模型。使用细胞计数试剂盒-8(CCK8)测定细胞存活率。实时定量逆转录聚合酶链反应(qRT-PCR)和蛋白质印迹分析用于检测microRNA-671-5p和双特异性磷酸酶(DUSP)8蛋白的表达,分别,白细胞介素（IL）-6，IL-1β，采用qRT-PCR和酶联免疫吸附试验(ELISA)检测肿瘤坏死因子(TNF)-α。使用双荧光素酶报告系统来确定micoRNA-671-5p与DUSP8表达之间的关系。使用蛋白质印迹分析证实p38丝裂原活化蛋白激酶(MAPK)信号通路的激活。用LPS处理hPDLSCs后，microRNA-671-5p在hPDLSCs中的表达水平升高,细胞活力下降，炎症因子的表达呈上升趋势。MicroRNA-671-5p靶向并结合DUSP8。沉默microRNA-671-5p或过表达DUSP8可以提高细胞存活率并减少炎症反应。当DUSP8过表达时，p-p38的表达降低。
结论：microRNA-671-5p靶向DUSP8/p38MAPK通路以调节LPS诱导的hPDLSCs增殖和炎症。
