Andrukhov Oleh, Ertlschweiger Sandra, Moritz Andreas, Bantleon Hans-Peter, Rausch-Fan Xiaohui
Central Research Unit, Bernhard Gottlieb University Clinic of Dentistry, Medical University of Vienna.
Acta Odontol Scand. 2014 Jul;72(5):337-45. doi: 10.3109/00016357.2013.834535. Epub 2013 Nov 21.
Gingival fibroblasts (GFs) produce pro-inflammatory cytokines in response to stimulation with lipopolysaccharide (LPS) of Porphyromonas gingivalis, which is thought to be mediated by activation of toll-like receptors (TLR)2 and TLR4. The present study investigated the expression of interleukin (IL)-6, TLR2, and TLR4 in GFs of seven different donors upon stimulation with P. gingivalis LPS. The effects of P. gingivalis LPS were compared with those of TLR4 agonist Escherichia coli LPS and TLR2 agonist Pam3CSK4.
GFs were stimulated with P. gingivalis LPS, E. coli LPS or Pam3CSK4 and the expression of IL-6, TLR2 and TLR4 was measured by qPCR. The surface expression of TLR2 and TLR4 was measured by flow cytometry.
In GFs from three donors, P. gingivalis LPS and Pam3CSK4 induced a markedly lower increase in IL-6 expression than E. coli LPS. This was accompanied by significant down-regulation of the TLR2 and TLR4 expression. In GFs from another four donors, an increase in IL-6 expression upon stimulation with P. gingivalis LPS and Pam3CSK4 was similar or even higher than that induced by E. coli LPS. In GFs of these donors, all stimuli induced an up-regulation of both mRNA and protein expression of TLR2 and did not influence that of TLR4.
This study suggests that P. gingivalis LPS and E. coli LPS differently regulate cytokine production in human gingival fibroblasts. Regulation of the expression level of TLR2 and TLR4 by periodontal pathogens might be an important factor controlling the inflammatory response in GFs.
牙龈成纤维细胞(GFs)在受到牙龈卟啉单胞菌脂多糖(LPS)刺激时会产生促炎细胞因子,这一过程被认为是由Toll样受体(TLR)2和TLR4的激活介导的。本研究调查了七种不同供体的牙龈成纤维细胞在受到牙龈卟啉单胞菌LPS刺激后白细胞介素(IL)-6、TLR2和TLR4的表达情况。将牙龈卟啉单胞菌LPS的作用与TLR4激动剂大肠杆菌LPS和TLR2激动剂Pam3CSK4的作用进行了比较。
用牙龈卟啉单胞菌LPS、大肠杆菌LPS或Pam3CSK4刺激牙龈成纤维细胞,通过qPCR检测IL-6、TLR2和TLR)4的表达。通过流式细胞术检测TLR2和TLR4的表面表达。
在来自三名供体的牙龈成纤维细胞中,牙龈卟啉单胞菌LPS和Pam3CSK4诱导的IL-6表达增加明显低于大肠杆菌LPS。这伴随着TLR2和TLR4表达的显著下调。在来自另外四名供体的牙龈成纤维细胞中,牙龈卟啉单胞菌LPS和Pam3CSK4刺激后IL-6表达增加与大肠杆菌LPS诱导的相似甚至更高。在这些供体的牙龈成纤维细胞中,所有刺激均诱导TLR2的mRNA和蛋白表达上调,且不影响TLR4的表达。
本研究表明,牙龈卟啉单胞菌LPS和大肠杆菌LPS对人牙龈成纤维细胞中细胞因子产生的调节方式不同。牙周病原体对TLR2和TLR4表达水平的调节可能是控制牙龈成纤维细胞炎症反应的一个重要因素。