Melatonin Receptor Agonists as the "Perioceutics" Agents for Periodontal Disease through Modulation of Porphyromonas gingivalis Virulence and Inflammatory Response.
Melatonin Receptor Agonists as the "Perioceutics" Agents for Periodontal Disease through Modulation of Porphyromonas gingivalis Virulence and Inflammatory Response.
复制标题
DOI：
10.1371/journal.pone.0166442
复制
发表时间：
2016
期刊：
影响因子：
3.7
通讯作者：
Song ZC
中科院分区：
文献类型：
--
作者：
Zhou W;Zhang X;Zhu CL;He ZY;Liang JP;Song ZC
牙周药物包括抗菌治疗和宿主调节治疗，已成为牙周病的重要治疗手段。牙周病患者的褪黑素水平明显降低，提示褪黑素可能作为牙周病的一种潜在的“牙周治疗剂”。本研究旨在探讨褪黑素受体激动剂（褪黑素和雷美替酮）对牙龈卟啉单胞菌毒力和牙龈卟啉单胞菌衍生脂多糖（Pg-LPS）诱导的炎症的影响。用微量稀释法测定褪黑素受体激动剂对牙龈卟啉单胞菌增殖培养物的影响。结晶紫染色法和MTT法分别检测牙龈卟啉单胞菌生物膜的形成、减少和活性。同时，用激光共聚焦扫描显微镜（CLSM）观察生物膜的形成。用发色肽和绵羊红细胞法测定了其对牙龈卟啉单胞菌牙龈蛋白酶和溶血活性的影响。采用RT-PCR方法检测毒力和铁/血红素利用率的mRNA表达。此外，通过MTT测定法评估褪黑激素受体激动剂对人牙龈成纤维细胞（HGF）的细胞活力。在用褪黑素受体激动剂预处理后，用Pg-LPS刺激HGF，然后通过酶联免疫吸附测定（ELISA）测量细胞因子（IL-6和IL-8）的释放。褪黑激素和雷美替酮确实表现出对培养物的抗菌作用。重要的是，它们抑制生物膜形成，减少已建立的生物膜，并降低牙龈卟啉单胞菌的生物膜活力。在亚最小抑菌浓度（sub-MIC）下，它们对牙龈卟啉菌蛋白酶的蛋白酶活性和溶血作用有明显的抑制作用，并呈剂量依赖性。它们在亚MIC浓度下显著抑制毒力因子（kgp、rgpA、rgpB、hagA和ragA）的mRNA表达，同时增加铁蛋白（ftn）或溶血素（hem）的mRNA表达。它们对HGF没有明显的细胞毒性。它们抑制Pg-LPS诱导的IL-6和IL-8的分泌，这被褪黑素受体拮抗剂luzindole逆转。褪黑激素受体激动剂可通过影响牙龈卟啉单胞菌的毒力特性以及Pg-LPS诱导的炎症反应来抑制牙龈卟啉单胞菌的增殖和生物膜生长。我们的研究提供了新的证据，褪黑激素受体激动剂可能是有用的新的“牙周”剂，以预防和治疗牙龈卟啉单胞菌相关的牙周疾病。
“Perioceutics” including antimicrobial therapy and host modulatory therapy has emerged as a vital adjunctive treatment of periodontal disease. Melatonin level was significantly reduced in patients with periodontal diseases suggesting melatonin could be applied as a potential “perioceutics” treatment of periodontal diseases. This study aims to investigate the effects of melatonin receptor agonists (melatonin and ramelteon) on Porphyromonas gingivalis virulence and Porphyromonas gingivalis-derived lipopolysaccharide (Pg-LPS)-induced inflammation. Effects of melatonin receptor agonists on Porphyromonas gingivalis planktonic cultures were determined by microplate dilution assays. Formation, reduction, and viability of Porphyromonas gingivalis biofilms were detected by crystal violet staining and MTT assays, respectively. Meanwhile, biofilms formation was also observed by confocal laser scanning microscopy (CLSM). The effects on gingipains and hemolytic activities of Porphyromonas gingivalis were evaluated using chromogenic peptides and sheep erythrocytes. The mRNA expression of virulence and iron/heme utilization was assessed using RT-PCR. In addition, cell viability of melatonin receptor agonists on human gingival fibroblasts (HGFs) was evaluated by MTT assays. After pretreatment of melatonin receptor agonists, HGFs were stimulated with Pg-LPS and then release of cytokines (IL-6 and lL-8) was measured by enzyme-linked immunosorbent assay (ELISA). Melatonin and ramelteon did exhibit antimicrobial effects against planktonic culture. Importantly, they inhibited biofilm formation, reduced the established biofilms, and decreased biofilm viability of Porphyromonas gingivalis. Furthermore, they at sub-minimum inhibitory concentration (sub-MIC) concentrations markedly inhibited the proteinase activities of gingipains and hemolysis in a dose-dependent manner. They at sub-MIC concentrations significantly inhibited the mRNA expression of virulence factors (kgp, rgpA, rgpB, hagA, and ragA), while increasing the mRNA expression of ferritin (ftn) or hemolysin (hem). They did not show obvious cytotoxicity toward HGFs. They inhibited Pg-LPS-induced IL-6 and IL-8 secretion, which was reversed by luzindole, the melatonin receptor antagonist. Melatonin receptor agonists can inhibit planktonic and biofilm growth of Porphyromonas gingivalis by affecting the virulent properties, as well as Pg-LPS-induced inflammatory response. Our study provides new evidence that melatonin receptor agonists might be useful as novel “perioceutics” agents to prevent and treat Porphyromonas gingivalis-associated periodontal diseases.
登录
查看更多内容
影响因子：
18.6
作者：
Jain S;Darveau RP
通讯作者：
Darveau RP
Keystone-Pathogen假设。
DOI：
10.1038/nrmicro2873
发表时间：
2012-10
期刊：
Nature reviews. Microbiology
影响因子：
--
作者：
通讯作者：
--
影响因子：
3.1
作者：
CHU, L;BRAMANTI, TE;HOLT, SC
通讯作者：
HOLT, SC
影响因子：
2.1
作者：
Chava VK;Sirisha K
通讯作者：
Sirisha K
影响因子：
10.3
作者：
Choi, Eun-Young;Jin, Ji-Young;Kim, Sung-Jo
通讯作者：
Kim, Sung-Jo