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人牙周韧带细胞在受到牙龈卟啉单胞菌脂多糖刺激时不会表现出内毒素耐受。

Human periodontal ligament cells exhibit no endotoxin tolerance upon stimulation with Porphyromonas gingivalis lipopolysaccharide.

机构信息

School of Dentistry, Medical University of Vienna, Vienna, Austria.

出版信息

J Periodontal Res. 2018 Aug;53(4):589-597. doi: 10.1111/jre.12549. Epub 2018 Mar 26.

Abstract

BACKGROUND/OBJECTIVES: Endotoxin tolerance is characterized by a state of hyporesponsiveness after confrontation with endotoxins such as lipopolysaccharides (LPS) at low concentrations. The aim of this study was to investigate, whether pretreatment with Porphyromonas gingivalis leads to endotoxin tolerance induction and possible alterations in toll-like receptor (TLR) 2- and 4-induced response in human periodontal ligament cells (hPDLCs).

MATERIAL AND METHODS

Primary hPDLCs were pretreated with P. gingivalis (0.1 or 0.3 μg/mL) LPS for 24 hours and afterwards treated with one of the following stimuli: P. gingivalis LPS (1 μg/mL); TLR4 agonist Escherichia coli LPS (0.1 μg/mL; 1 μg/mL); TLR2 agonist Pam3CSK4 (0.1 μg/mL; 1 μg/mL). The protein expression of interleukin (IL)-6, IL-8 and monocyte chemotactic protein-1 was analyzed with quantitative polymerase chain reaction and enzyme-linked immunosorbent assay. Gene expression levels of TLR2 and TLR4 were determined by quantitative polymerase chain reaction.

RESULTS

Pretreatment of cells with low concentrations of P. gingivalis LPS did not result in lower production of IL-6, IL-8 and monocyte chemotactic protein-1 compared to control group. In some cases, pretreated cells exhibited lower gene expression levels of TLR2 and TLR4 compared to non-pretreated cells.

CONCLUSION

The results of this study implicate that hPDLCs do not develop endotoxin tolerance. Furthermore, the amplitude of the inflammatory response shows no significant dependency on TLR2 and TLR4 expression levels.

摘要

背景/目的:内毒素耐受是指在低浓度内毒素(如脂多糖[LPS])作用下,机体产生的一种低反应状态。本研究旨在探讨牙龈卟啉单胞菌预处理是否会导致内毒素耐受诱导,并可能改变人牙周膜细胞(hPDLC)中 Toll 样受体(TLR)2 和 TLR4 诱导的反应。

材料和方法

用牙龈卟啉单胞菌 LPS(0.1 或 0.3μg/mL)预处理原代 hPDLC 24 小时,然后用以下刺激物之一进行处理:牙龈卟啉单胞菌 LPS(1μg/mL);TLR4 激动剂大肠杆菌 LPS(0.1μg/mL;1μg/mL);TLR2 激动剂 Pam3CSK4(0.1μg/mL;1μg/mL)。用定量聚合酶链反应和酶联免疫吸附试验分析白细胞介素(IL)-6、IL-8 和单核细胞趋化蛋白-1 的蛋白表达。用定量聚合酶链反应测定 TLR2 和 TLR4 的基因表达水平。

结果

与对照组相比,用低浓度牙龈卟啉单胞菌 LPS 预处理细胞不会导致 IL-6、IL-8 和单核细胞趋化蛋白-1 的产生减少。在某些情况下,与未经预处理的细胞相比,预处理细胞的 TLR2 和 TLR4 基因表达水平较低。

结论

本研究结果表明,hPDLC 不会产生内毒素耐受。此外,炎症反应的幅度与 TLR2 和 TLR4 的表达水平无明显相关性。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/1d47/6055822/e234e8fa1f70/JRE-53-589-g001.jpg

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